Antagonists of the kappa opioid receptor

ABSTRACT

The present technology is directed to compounds, compositions, and methods related to non-morphinan-like kappa opioid receptor (KOR) antagonists. The technology is suited to treat addiction, diuresis, depression, post traumatic stress disorder, an eating disorder, panic disorder, social anxiety disorder, general anxiety disorder, obsessive compulsive disorders, excessive or unreasonable specific phobias, and/or other conditions related to anxiety or aversion-reward responses.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a U.S. national phase application under 35 U.S.C. §371 of International Application No. PCT/US2015/062708, filed on Nov.25, 2015, which claims the benefit of and priority to U.S. ProvisionalPatent Application No. 62/084,932, filed Nov. 26, 2014, the entiredisclosures of which are hereby incorporated by reference in theirentireties for any and all purposes.

GOVERNMENT RIGHTS

This invention was made with government support under Grant NumberR01-DA031927 awarded by the National Institutes of Health. Thegovernment has certain rights in the invention.

FIELD

The present technology is directed to compounds, compositions, andmethods related to non-morphinan-like kappa opioid receptor (KOR)antagonists. The technology is suited to treat addiction, diuresis,depression, post-traumatic stress disorder, an eating disorder, panicdisorder, social anxiety disorder, general anxiety disorder, obsessivecompulsive disorders, excessive or unreasonable specific phobias, and/orother conditions related to anxiety or aversion-reward responses.

SUMMARY

In an aspect, a compound according to formula I is provided

where G¹ and G² are each independently C═O or S(O)₂; R¹, R², R³, R⁴, R⁵,R⁷, R⁸, R⁹, R¹⁰, and R¹¹ are each independently H, halo, hydroxy, amino,cyano, trifluoromethyl, thiol, alkylthio, sulfoxide, sulfone, nitro,pentafluorosulfanyl, carboxylate, amide, ester, or a substituted orunsubstituted C₁-C₆ alkyl, C₁-C₆ alkoxy, aryl, aryloxy, C₁-C₆ alkanoyl,C₁-C₈ alkanoyloxy, aryloyl, or aryloyloxy group, where any two adjacentR¹, R², R³, R⁴, R⁵, R⁷, R⁸, R⁹, R¹⁰, and R¹¹ may join to form a5-membered or 6-membered substituted or unsubstituted heteroalkyl group;R⁶ is a branched C₁-C₈ alkyl group or a substituted or unsubstitutedcycloalkyl or aryl group; R¹² and R¹³ are each independently H or asubstituted or unsubstituted C₁-C₈ alkyl or C₅-C₇ cycloalkyl group; andn is 0, 1, or 2; or stereoisomers, tautomers, solvates, and/or saltsthereof; provided that when G¹ is S(O)₂, G² is C═O, R¹, R², R⁴, R⁵, R⁷,R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³ are each H, R⁶ is isopropyl, and n is 1, thenR³ is not methyl.

In a related aspect, a composition is provided that includes thecompound of any one of the above embodiments and a pharmaceuticallyacceptable carrier.

In another aspect, a pharmaceutical composition is provided, thepharmaceutical composition including an effective amount of the compoundof any one of the above embodiments for treating a condition, whereinthe condition is addiction, diuresis, depression, post-traumatic stressdisorder, an eating disorder, panic disorder, social anxiety disorder,general anxiety disorder, obsessive compulsive disorders, excessive orunreasonable specific phobias, and/or other conditions related toanxiety or aversion-reward responses.

In another aspect, a method is provided that includes administering aneffective amount of a compound of any one of the above embodiments, oradministering a pharmaceutical composition including an effective amountof a compound of any one of the above embodiments, to a subjectsuffering from addiction, diuresis, depression, post-traumatic stressdisorder, an eating disorder, panic disorder, social anxiety disorder,general anxiety disorder, obsessive compulsive disorders, excessive orunreasonable specific phobias, and/or other conditions related toanxiety or aversion-reward responses.

In another aspect, a method is provided that includes inhibitingβ-arrestin recruitment in a subject by administering an effective amountof a compound of any one of the above embodiments. The subject may besuffering from addiction, diuresis, depression, post-traumatic stressdisorder, an eating disorder, panic disorder, social anxiety disorder,general anxiety disorder, obsessive compulsive disorders, excessive orunreasonable specific phobias, and/or other conditions related toanxiety or aversion-reward responses. In any embodiment herein, thesubject may be suffering from addiction. The subject may be sufferingfrom addiction to at least one of nicotine, ethanol, cocaine, opioids,amphetamines, marijuana, and synthetic cannabinoid agonists. The methodmay include inhibiting β-arrestin2 recruitment.

In another aspect, a method is provided for treating an addiction in asubject where the method includes administering an effective amount of acompound of any one of the above embodiments. In some embodiments, theaddiction is to at least one of nicotine, ethanol, cocaine, opioids,amphetamines, marijuana, and synthetic cannabinoid agonists.

In an aspect, method of inhibiting β-arrestin recruitment is provided,where the method includes contacting a kappa opioid receptor with acompound of any one of the above embodiments. The method may includeinhibiting β-arrestin2 recruitment.

DESCRIPTION OF THE DRAWINGS

FIGS. 1A-D shows the results of experiments comparing the detectedconcentrations of a compound of the present technology with amorphinan-like opioid antagonist, norBNI, in the plasma and brain tissueof mice over the time period necessary for clearance (or up to 72 h)following a single 10 mg/kg IP dose. FIGS. 1A and 1B show theconcentration of a compound of the present technology and norBNI(respectively) in brain tissue, and FIGS. 1C and 1D show theconcentration of a compound of the present technology and norBNI(respectively) in plasma.

FIG. 2 shows the locomotor activity in mice for one embodiment of acompound of the present technology compared to known kappa opiod agonistU50, 488. The animal numbers are shown in parentheses.

FIG. 3 shows the locomotor activity in mice pretreated with oneembodiment of a compound of the present technology followed by treatmentwith U50,488 in comparison to mice that received a vehicle pretreatmentfollowed by U50,488. The animal numbers are shown in parentheses.

DETAILED DESCRIPTION

In various aspects, the present technology provides compounds andmethods for antagonizing a kappa opioid receptor. The compounds providedherein can be formulated into pharmaceutical compositions andmedicaments that are useful in the disclosed methods. Also provided isthe use of the compounds in preparing pharmaceutical formulations andmedicaments.

The following terms are used throughout as defined below.

As used herein and in the appended claims, singular articles such as “a”and “an” and “the” and similar referents in the context of describingthe elements (especially in the context of the following claims) are tobe construed to cover both the singular and the plural, unless otherwiseindicated herein or clearly contradicted by context. Recitation ofranges of values herein are merely intended to serve as a shorthandmethod of referring individually to each separate value falling withinthe range, unless otherwise indicated herein, and each separate value isincorporated into the specification as if it were individually recitedherein. All methods described herein can be performed in any suitableorder unless otherwise indicated herein or otherwise clearlycontradicted by context. The use of any and all examples, or exemplarylanguage (e.g., “such as”) provided herein, is intended merely to betterilluminate the embodiments and does not pose a limitation on the scopeof the claims unless otherwise stated. No language in the specificationshould be construed as indicating any non-claimed element as essential.

As used herein, “about” will be understood by persons of ordinary skillin the art and will vary to some extent depending upon the context inwhich it is used. If there are uses of the term which are not clear topersons of ordinary skill in the art, given the context in which it isused, “about” will mean up to plus or minus 10% of the particular term.

Generally, reference to a certain element such as hydrogen or H is meantto include all isotopes of that element. For example, if an R group isdefined to include hydrogen or H, it also includes deuterium andtritium. Compounds comprising radioisotopes such as tritium, C¹⁴, P³²and S³⁵ are thus within the scope of the present technology. Proceduresfor inserting such labels into the compounds of the present technologywill be readily apparent to those skilled in the art based on thedisclosure herein.

In general, “substituted” refers to an organic group as defined below(e.g., an alkyl group) in which one or more bonds to a hydrogen atomcontained therein are replaced by a bond to non-hydrogen or non-carbonatoms. Substituted groups also include groups in which one or more bondsto a carbon(s) or hydrogen(s) atom are replaced by one or more bonds,including double or triple bonds, to a heteroatom. Thus, a substitutedgroup is substituted with one or more substituents, unless otherwisespecified. In some embodiments, a substituted group is substituted with1, 2, 3, 4, 5, or 6 substituents. Examples of substituent groupsinclude: halogens (i.e., F, Cl, Br, and I); hydroxyls; alkoxy, alkenoxy,aryloxy, aralkyloxy, heterocyclyl, heterocyclylalkyl, heterocyclyloxy,and heterocyclylalkoxy groups; carbonyls (oxo); carboxylates; esters;urethanes; oximes; hydroxylamines; alkoxyamines; aralkoxyamines; thiols;sulfides; sulfoxides; sulfones; sulfonyls; pentafluorosulfanyl (i.e.,SF₅), sulfonamides; amines; N-oxides; hydrazines; hydrazides;hydrazones; azides; amides; ureas; amidines; guanidines; enamines;imides; isocyanates; isothiocyanates; cyanates; thiocyanates; imines;nitro groups; nitriles (i.e., CN); and the like.

Substituted ring groups such as substituted cycloalkyl, aryl,heterocyclyl and heteroaryl groups also include rings and ring systemsin which a bond to a hydrogen atom is replaced with a bond to a carbonatom. Therefore, substituted cycloalkyl, aryl, heterocyclyl andheteroaryl groups may also be substituted with substituted orunsubstituted alkyl, alkenyl, and alkynyl groups as defined below.

Alkyl groups include straight chain and branched chain alkyl groupshaving from 1 to 12 carbon atoms, and typically from 1 to 10 carbons or,in some embodiments, from 1 to 8, 1 to 6, or 1 to 4 carbon atoms.Examples of straight chain alkyl groups include groups such as methyl,ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octylgroups. Examples of branched alkyl groups include, but are not limitedto, isopropyl, iso-butyl, sec-butyl, tert-butyl, neopentyl, isopentyl,and 2,2-dimethylpropyl groups. Representative substituted alkyl groupsmay be substituted one or more times with substituents such as thoselisted above, and include without limitation haloalkyl (e.g.,trifluoromethyl), hydroxyalkyl, thioalkyl, aminoalkyl, alkylaminoalkyl,dialkylaminoalkyl, alkoxyalkyl, carboxyalkyl, and the like.

Cycloalkyl groups include mono-, bi- or tricyclic alkyl groups havingfrom 3 to 12 carbon atoms in the ring(s), or, in some embodiments, 3 to10, 3 to 8, or 3 to 4, 5, or 6 carbon atoms. Exemplary monocycliccycloalkyl groups include, but not limited to, cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl groups. In someembodiments, the cycloalkyl group has 3 to 8 ring members, whereas inother embodiments the number of ring carbon atoms range from 3 to 5, 3to 6, or 3 to 7. Bi- and tricyclic ring systems include both bridgedcycloalkyl groups and fused rings, such as, but not limited to,bicyclo[2.1.1]hexane, adamantyl, decalinyl, and the like. Substitutedcycloalkyl groups may be substituted one or more times with,non-hydrogen and non-carbon groups as defined above. However,substituted cycloalkyl groups also include rings that are substitutedwith straight or branched chain alkyl groups as defined above.Representative substituted cycloalkyl groups may be mono-substituted orsubstituted more than once, such as, but not limited to, 2,2-, 2,3-,2,4- 2,5- or 2,6-disubstituted cyclohexyl groups, which may besubstituted with substituents such as those listed above.

Cycloalkylalkyl groups are alkyl groups as defined above in which ahydrogen or carbon bond of an alkyl group is replaced with a bond to acycloalkyl group as defined above. In some embodiments, cycloalkylalkylgroups have from 4 to 16 carbon atoms, 4 to 12 carbon atoms, andtypically 4 to 10 carbon atoms. Substituted cycloalkylalkyl groups maybe substituted at the alkyl, the cycloalkyl or both the alkyl andcycloalkyl portions of the group. Representative substitutedcycloalkylalkyl groups may be mono-substituted or substituted more thanonce, such as, but not limited to, mono-, di- or tri-substituted withsubstituents such as those listed above.

Alkenyl groups include straight and branched chain alkyl groups asdefined above, except that at least one double bond exists between twocarbon atoms. Alkenyl groups have from 2 to 12 carbon atoms, andtypically from 2 to 10 carbons or, in some embodiments, from 2 to 8, 2to 6, or 2 to 4 carbon atoms. In some embodiments, the alkenyl group hasone, two, or three carbon-carbon double bonds. Examples include, but arenot limited to vinyl, allyl, —CH═CH(CH₃), —CH═C(CH₃)₂, —C(CH₃)═CH₂,—C(CH₃)═CH(CH₃), —C(CH₂CH₃)═CH₂, among others. Representativesubstituted alkenyl groups may be mono-substituted or substituted morethan once, such as, but not limited to, mono-, di- or tri-substitutedwith substituents such as those listed above.

Cycloalkenyl groups include cycloalkyl groups as defined above, havingat least one double bond between two carbon atoms. In some embodimentsthe cycloalkenyl group may have one, two or three double bonds but doesnot include aromatic compounds. Cycloalkenyl groups have from 4 to 14carbon atoms, or, in some embodiments, 5 to 14 carbon atoms, 5 to 10carbon atoms, or even 5, 6, 7, or 8 carbon atoms. Examples ofcycloalkenyl groups include cyclohexenyl, cyclopentenyl,cyclohexadienyl, cyclobutadienyl, and cyclopentadienyl.

Cycloalkenylalkyl groups are alkyl groups as defined above in which ahydrogen or carbon bond of the alkyl group is replaced with a bond to acycloalkenyl group as defined above. Substituted cycloalkenylalkylgroups may be substituted at the alkyl, the cycloalkenyl or both thealkyl and cycloalkenyl portions of the group. Representative substitutedcycloalkenylalkyl groups may be substituted one or more times withsubstituents such as those listed above.

Alkynyl groups include straight and branched chain alkyl groups asdefined above, except that at least one triple bond exists between twocarbon atoms. Alkynyl groups have from 2 to 12 carbon atoms, andtypically from 2 to 10 carbons or, in some embodiments, from 2 to 8, 2to 6, or 2 to 4 carbon atoms. In some embodiments, the alkynyl group hasone, two, or three carbon-carbon triple bonds. Examples include, but arenot limited to —C≡CH, —C≡CCH₃, —CH₂C≡CCH₃, —C≡CCH₂CH(CH₂CH₃)₂, amongothers. Representative substituted alkynyl groups may bemono-substituted or substituted more than once, such as, but not limitedto, mono-, di- or tri-substituted with substituents such as those listedabove.

Aryl groups are cyclic aromatic hydrocarbons that do not containheteroatoms. Aryl groups herein include monocyclic, bicyclic andtricyclic ring systems. Thus, aryl groups include, but are not limitedto, phenyl, azulenyl, heptalenyl, biphenyl, fluorenyl, phenanthrenyl,anthracenyl, indenyl, indanyl, pentalenyl, and naphthyl groups. In someembodiments, aryl groups contain 6-14 carbons, and in others from 6 to12 or even 6-10 carbon atoms in the ring portions of the groups. In someembodiments, the aryl groups are phenyl or naphthyl. Although the phrase“aryl groups” includes groups containing fused rings, such as fusedaromatic-aliphatic ring systems (e.g., indanyl, tetrahydronaphthyl, andthe like), it does not include aryl groups that have other groups, suchas alkyl or halo groups, bonded to one of the ring members. Rather,groups such as tolyl are referred to as substituted aryl groups.Representative substituted aryl groups may be mono-substituted orsubstituted more than once. For example, monosubstituted aryl groupsinclude, but are not limited to, 2-, 3-, 4-, 5-, or 6-substituted phenylor naphthyl groups, which may be substituted with substituents such asthose listed above.

Aralkyl groups are alkyl groups as defined above in which a hydrogen orcarbon bond of an alkyl group is replaced with a bond to an aryl groupas defined above. In some embodiments, aralkyl groups contain 7 to 16carbon atoms, 7 to 14 carbon atoms, or 7 to 10 carbon atoms. Substitutedaralkyl groups may be substituted at the alkyl, the aryl or both thealkyl and aryl portions of the group. Representative aralkyl groupsinclude but are not limited to benzyl and phenethyl groups and fused(cycloalkylaryl)alkyl groups such as 4-indanylethyl. Representativesubstituted aralkyl groups may be substituted one or more times withsubstituents such as those listed above.

Heterocyclyl groups include aromatic (also referred to as heteroaryl)and non-aromatic ring compounds containing 3 or more ring members, ofwhich one or more is a heteroatom such as, but not limited to, N, O, andS. In some embodiments, the heterocyclyl group contains 1, 2, 3 or 4heteroatoms. In some embodiments, heterocyclyl groups include mono-, bi-and tricyclic rings having 3 to 16 ring members, whereas other suchgroups have 3 to 6, 3 to 10, 3 to 12, or 3 to 14 ring members.Heterocyclyl groups encompass aromatic, partially unsaturated andsaturated ring systems, such as, for example, imidazolyl, imidazolinyland imidazolidinyl groups. The phrase “heterocyclyl group” includesfused ring species including those comprising fused aromatic andnon-aromatic groups, such as, for example, benzotriazolyl,2,3-dihydrobenzo[1,4]dioxinyl, and benzo[1,3]dioxolyl. The phrase alsoincludes bridged polycyclic ring systems containing a heteroatom suchas, but not limited to, quinuclidyl. However, the phrase does notinclude heterocyclyl groups that have other groups, such as alkyl, oxoor halo groups, bonded to one of the ring members. Rather, these arereferred to as “substituted heterocyclyl groups”. Heterocyclyl groupsinclude, but are not limited to, aziridinyl, azetidinyl, pyrrolidinyl,imidazolidinyl, pyrazolidinyl, thiazolidinyl, tetrahydrothiophenyl,tetrahydrofuranyl, dioxolyl, furanyl, thiophenyl, pyrrolyl, pyrrolinyl,imidazolyl, imidazolinyl, pyrazolyl, pyrazolinyl, triazolyl, tetrazolyl,oxazolyl, isoxazolyl, thiazolyl, thiazolinyl, isothiazolyl,thiadiazolyl, oxadiazolyl, piperidyl, piperazinyl, morpholinyl,thiomorpholinyl, tetrahydropyranyl, tetrahydrothiopyranyl, oxathiane,dioxyl, dithianyl, pyranyl, pyridyl, pyrimidinyl, pyridazinyl,pyrazinyl, triazinyl, dihydropyridyl, dihydrodithiinyl,dihydrodithionyl, homopiperazinyl, quinuclidyl, indolyl, indolinyl,isoindolyl, azaindolyl (pyrrolopyridyl), indazolyl, indolizinyl,benzotriazolyl, benzimidazolyl, benzofuranyl, benzothiophenyl,benzthiazolyl, benzoxadiazolyl, benzoxazinyl, benzodithiinyl,benzoxathiinyl, benzothiazinyl, benzoxazolyl, benzothiazolyl,benzothiadiazolyl, benzo[1,3]dioxolyl, pyrazolopyridyl, imidazopyridyl(azabenzimidazolyl), triazolopyridyl, isoxazolopyridyl, purinyl,xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl, quinolizinyl,quinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl, naphthyridinyl,pteridinyl, thianaphthyl, dihydrobenzothiazinyl, dihydrobenzofuranyl,dihydroindolyl, dihydrobenzodioxinyl, tetrahydroindolyl,tetrahydroindazolyl, tetrahydrobenzimidazolyl, tetrahydrobenzotriazolyl,tetrahydropyrrolopyridyl, tetrahydropyrazolopyridyl,tetrahydroimidazopyridyl, tetrahydrotriazolopyridyl, andtetrahydroquinolinyl groups. Representative substituted heterocyclylgroups may be mono-substituted or substituted more than once, such as,but not limited to, pyridyl or morpholinyl groups, which are 2-, 3-, 4-,5-, or 6-substituted, or disubstituted with various substituents such asthose listed above.

Heteroaryl groups are aromatic ring compounds containing 5 or more ringmembers, of which, one or more is a heteroatom such as, but not limitedto, N, O, and S. Heteroaryl groups include, but are not limited to,groups such as pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl,isoxazolyl, thiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl,thiophenyl, benzothiophenyl, furanyl, benzofuranyl, indolyl, azaindolyl(pyrrolopyridinyl), indazolyl, benzimidazolyl, imidazopyridinyl(azabenzimidazolyl), pyrazolopyridinyl, triazolopyridinyl,benzotriazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl,imidazopyridinyl, isoxazolopyridinyl, thianaphthyl, purinyl, xanthinyl,adeninyl, guaninyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl,quinoxalinyl, and quinazolinyl groups. Heteroaryl groups include fusedring compounds in which all rings are aromatic such as indolyl groupsand include fused ring compounds in which only one of the rings isaromatic, such as 2,3-dihydro indolyl groups. Although the phrase“heteroaryl groups” includes fused ring compounds, the phrase does notinclude heteroaryl groups that have other groups bonded to one of thering members, such as alkyl groups. Rather, heteroaryl groups with suchsubstitution are referred to as “substituted heteroaryl groups.”Representative substituted heteroaryl groups may be substituted one ormore times with various substituents such as those listed above.

Heterocyclylalkyl groups are alkyl groups as defined above in which ahydrogen or carbon bond of an alkyl group is replaced with a bond to aheterocyclyl group as defined above. Substituted heterocyclylalkylgroups may be substituted at the alkyl, the heterocyclyl or both thealkyl and heterocyclyl portions of the group. Representativeheterocyclyl alkyl groups include, but are not limited to,morpholin-4-yl-ethyl, furan-2-yl-methyl, imidazol-4-yl-methyl,pyridin-3-yl-methyl, tetrahydrofuran-2-yl-ethyl, and indol-2-yl-propyl.Representative substituted heterocyclylalkyl groups may be substitutedone or more times with substituents such as those listed above.

Heteroaralkyl groups are alkyl groups as defined above in which ahydrogen or carbon bond of an alkyl group is replaced with a bond to aheteroaryl group as defined above. Substituted heteroaralkyl groups maybe substituted at the alkyl, the heteroaryl or both the alkyl andheteroaryl portions of the group. Representative substitutedheteroaralkyl groups may be substituted one or more times withsubstituents such as those listed above.

Groups described herein having two or more points of attachment (i.e.,divalent, trivalent, or polyvalent) within the compound of the presenttechnology are designated by use of the suffix, “ene.” For example,divalent alkyl groups are alkylene groups, divalent aryl groups arearylene groups, divalent heteroaryl groups are divalent heteroarylenegroups, and so forth. Substituted groups having a single point ofattachment to the compound of the present technology are not referred tousing the “ene” designation. Thus, e.g., chloroethyl is not referred toherein as chloroethylene.

Alkoxy groups are hydroxyl groups (—OH) in which the bond to thehydrogen atom is replaced by a bond to a carbon atom of a substituted orunsubstituted alkyl group as defined above. Examples of linear alkoxygroups include but are not limited to methoxy, ethoxy, propoxy, butoxy,pentoxy, hexoxy, and the like. Examples of branched alkoxy groupsinclude but are not limited to isopropoxy, sec-butoxy, tert-butoxy,isopentoxy, isohexoxy, and the like. Examples of cycloalkoxy groupsinclude but are not limited to cyclopropyloxy, cyclobutyloxy,cyclopentyloxy, cyclohexyloxy, and the like. Representative substitutedalkoxy groups may be substituted one or more times with substituentssuch as those listed above.

The terms “alkanoyl” and “alkanoyloxy” as used herein can refer,respectively, to —C(O)-alkyl groups and —O—C(O)-alkyl groups, eachcontaining 2-5 carbon atoms. Similarly, “aryloyl” and “aryloyloxy” referto —C(O)-aryl groups and —O—C(O)-aryl groups.

The terms “aryloxy” and “arylalkoxy” refer to, respectively, asubstituted or unsubstituted aryl group bonded to an oxygen atom and asubstituted or unsubstituted aralkyl group bonded to the oxygen atom atthe alkyl. Examples include but are not limited to phenoxy, naphthyloxy,and benzyloxy. Representative substituted aryloxy and arylalkoxy groupsmay be substituted one or more times with substituents such as thoselisted above.

The term “carboxylate” as used herein refers to a —COOH group.

The term “ester” as used herein refers to —COOR⁷⁰ and —C(O)O-G groups.R⁷⁰ is a substituted or unsubstituted alkyl, cycloalkyl, alkenyl,alkynyl, aryl, aralkyl, heterocyclylalkyl or heterocyclyl group asdefined herein. G is a carboxylate protecting group. Carboxylateprotecting groups are well known to one of ordinary skill in the art. Anextensive list of protecting groups for the carboxylate groupfunctionality may be found in Protective Groups in Organic Synthesis,Greene, T. W.; Wuts, P. G. M., John Wiley & Sons, New York, N.Y., (3rdEdition, 1999) which can be added or removed using the procedures setforth therein and which is hereby incorporated by reference in itsentirety and for any and all purposes as if fully set forth herein.

The term “amide” (or “amido”) includes C- and N-amide groups, i.e.,—C(O)NR⁷¹R⁷², and —NR⁷¹C(O)R⁷² groups, respectively. R⁷¹ and R⁷² areindependently hydrogen, or a substituted or unsubstituted alkyl,alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heterocyclylalkyl orheterocyclyl group as defined herein. Amido groups therefore include butare not limited to carbamoyl groups (—C(O)NH₂) and formamide groups(—NHC(O)H). In some embodiments, the amide is —NR⁷¹C(O)—(C₁₋₅ alkyl) andthe group is termed “carbonylamino,” and in others the amide is—NHC(O)-alkyl and the group is termed “alkanoylamino.”

The term “nitrile” or “cyano” as used herein refers to the —CN group.

Urethane groups include N- and O-urethane groups, i.e., —NR⁷³C(O)OR⁷⁴and —OC(O)NR⁷³R⁷⁴ groups, respectively. R⁷³ and R⁷⁴ are independently asubstituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, aryl,aralkyl, heterocyclylalkyl, or heterocyclyl group as defined herein. R⁷³may also be H.

The term “amine” (or “amino”) as used herein refers to —NR⁷⁵R⁷⁶ groups,wherein R⁷⁵ and R⁷⁶ are independently hydrogen, or a substituted orunsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl,heterocyclylalkyl or heterocyclyl group as defined herein. In someembodiments, the amine is alkylamino, dialkylamino, arylamino, oralkylarylamino. In other embodiments, the amine is NH₂, methylamino,dimethylamino, ethylamino, diethylamino, propylamino, isopropylamino,phenylamino, or benzylamino.

The term “sulfonamido” includes S- and N-sulfonamide groups, i.e.,—SO₂NR⁷⁸R⁷⁹ and —NR⁷⁸SO₂R⁷⁹ groups, respectively. R⁷⁸ and R⁷⁹ areindependently hydrogen, or a substituted or unsubstituted alkyl,alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heterocyclylalkyl, orheterocyclyl group as defined herein. Sulfonamido groups thereforeinclude but are not limited to sulfamoyl groups (—SO₂NH₂). In someembodiments herein, the sulfonamido is —NHSO₂-alkyl and is referred toas the “alkylsulfonylamino” group.

The term “thiol” refers to —SH groups, while “sulfides” include —SR⁸⁰groups, “sulfoxides” include —S(O)R⁸¹ groups, “sulfones” include —SO₂R⁸²groups, and “sulfonyls” include —SO₂OR⁸³. R⁸⁰, R⁸¹, R⁸², and R⁸³ areeach independently a substituted or unsubstituted alkyl, cycloalkyl,alkenyl, alkynyl, aryl aralkyl, heterocyclyl or heterocyclylalkyl groupas defined herein. In some embodiments the sulfide is an alkylthiogroup, —S-alkyl.

The term “urea” refers to —NR⁸⁴—C(O)—NR⁸⁵R⁸⁶ groups. R⁸⁴, R⁸⁵, and R⁸⁶groups are independently hydrogen, or a substituted or unsubstitutedalkyl, alkenyl, alkynyl, cycloalkyl, aryl, aralkyl, heterocyclyl, orheterocyclylalkyl group as defined herein.

The term “amidine” refers to —C(NR⁸⁷)NR⁸⁸R⁸⁹ and —NR⁸⁷C(NR⁸⁸)R⁸⁹,wherein R⁸⁷, R⁸⁸, and R⁸⁹ are each independently hydrogen, or asubstituted or unsubstituted alkyl, cycloalkyl, alkenyl, alkynyl, arylaralkyl, heterocyclyl or heterocyclylalkyl group as defined herein.

The term “guanidine” refers to —NR⁹⁰C(NR⁹¹)NR⁹²R⁹³, wherein R⁹⁰, R⁹¹,R⁹² and R⁹³ are each independently hydrogen, or a substituted orunsubstituted alkyl, cycloalkyl, alkenyl, alkynyl, aryl aralkyl,heterocyclyl or heterocyclylalkyl group as defined herein.

The term “enamine” refers to —C(R⁹⁴)═C(R⁹⁵)NR⁹⁶R⁹⁷ and—NR⁹⁴C(R⁹⁵)═C(R⁹⁶)R⁹⁷, wherein R⁹⁴, R⁹⁵, R⁹⁶ and R⁹⁷ are eachindependently hydrogen, a substituted or unsubstituted alkyl,cycloalkyl, alkenyl, alkynyl, aryl aralkyl, heterocyclyl orheterocyclylalkyl group as defined herein.

The term “halogen” or “halo” as used herein refers to bromine, chlorine,fluorine, or iodine. In some embodiments, the halogen is fluorine. Inother embodiments, the halogen is chlorine or bromine.

The term “hydroxyl” as used herein can refer to —OH or its ionized form,—O—. A “hydroxyalkyl” group is a hydroxyl-substituted alkyl group, suchas HO—CH₂—.

The term “imide” refers to —C(O)NR⁹⁸C(O)R⁹⁹, wherein R⁹⁸ and R⁹⁹ areeach independently hydrogen, or a substituted or unsubstituted alkyl,cycloalkyl, alkenyl, alkynyl, aryl aralkyl, heterocyclyl orheterocyclylalkyl group as defined herein.

The term “imine” refers to —CR¹⁰⁰(NR¹⁰¹) and —N(CR¹⁰⁰R¹⁰¹) groups,wherein R¹⁰⁰ and R¹⁰¹ are each independently hydrogen or a substitutedor unsubstituted alkyl, cycloalkyl, alkenyl, alkynyl, aryl aralkyl,heterocyclyl or heterocyclylalkyl group as defined herein, with theproviso that R¹⁰⁰ and R¹⁰¹ are not both simultaneously hydrogen.

The term “nitro” as used herein refers to an —NO₂ group.

The term “trifluoromethyl” as used herein refers to —CF₃.

The term “trifluoromethoxy” as used herein refers to —OCF₃.

The term “azido” refers to —N₃.

The term “trialkyl ammonium” refers to a —N(alkyl)₃ group. Atrialkylammonium group is positively charged and thus typically has anassociated anion, such as halogen anion.

The term “isocyano” refers to —NC.

The term “isothiocyano” refers to —NCS.

The term “pentafluorosulfanyl” refers to —SF₅.

The term “addictive substance” refers to those substances that wheninternalized can generate a compulsive desire and/or need for theaddictive substance that is habit forming. Without being bound bytheory, addictive substances activates the reward pathways of the brainof a subject in some manner, leading to a desire to repeat theinternalization of the addictive substance. Exemplary addictivesubstances include, but are not limited to, nicotine, ethanol, cocaine,opioids, amphetamines, marijuana, and synthetic cannabinoid agonists.

The phrase “selectively inhibits” as used herein will be understood bypersons of ordinary skill in the art and will vary to some extentdepending upon the context in which the phrase is used. If there areuses of the phrase which are not clear to persons of ordinary skill inthe art, given the context in which the phrase is used, the phrase atminimum refers to the compounds acting through a specific mechanism ofaction, resulting in fewer off-target effects because the compoundstarget a particular receptor over other receptors, such as a kappaopioid receptor over a g opioid receptor (MOR) and/or a δ opioidreceptor (DOR), and or because the compounds target a particularmechanism such as β-arrestin recruitment and/or β-arrestin2 recruitmentover other mechanisms. The phrase may further be modified as discussedherein.

As will be understood by one skilled in the art, for any and allpurposes, particularly in terms of providing a written description, allranges disclosed herein also encompass any and all possible subrangesand combinations of subranges thereof. Any listed range can be easilyrecognized as sufficiently describing and enabling the same range beingbroken down into at least equal halves, thirds, quarters, fifths,tenths, etc. As a non-limiting example, each range discussed herein canbe readily broken down into a lower third, middle third and upper third,etc. As will also be understood by one skilled in the art all languagesuch as “up to,” “at least,” “greater than,” “less than,” and the likeinclude the number recited and refer to ranges which can be subsequentlybroken down into subranges as discussed above. Finally, as will beunderstood by one skilled in the art, a range includes each individualmember. Thus, for example, a group having 1-3 atoms refers to groupshaving 1, 2, or 3 atoms. Similarly, a group having 1-5 atoms refers togroups having 1, 2, 3, 4, or 5 atoms, and so forth.

Pharmaceutically acceptable salts of compounds described herein arewithin the scope of the present technology and include acid or baseaddition salts which retain the desired pharmacological activity and isnot biologically undesirable (e.g., the salt is not unduly toxic,allergenic, or irritating, and is bioavailable). When the compound ofthe present technology has a basic group, such as, for example, an aminogroup, pharmaceutically acceptable salts can be formed with inorganicacids (such as hydrochloric acid, hydroboric acid, nitric acid, sulfuricacid, and phosphoric acid), organic acids (e.g. alginate, formic acid,acetic acid, benzoic acid, gluconic acid, fumaric acid, oxalic acid,tartaric acid, lactic acid, maleic acid, citric acid, succinic acid,malic acid, methanesulfonic acid, benzenesulfonic acid, naphthalenesulfonic acid, and p-toluenesulfonic acid) or acidic amino acids (suchas aspartic acid and glutamic acid). When the compound of the presenttechnology has an acidic group, such as for example, a carboxylic acidgroup, it can form salts with metals, such as alkali and earth alkalimetals (e.g. Na⁺, Li⁺, K⁺, Ca²⁺, Mg²⁺, Zn²⁺), ammonia or organic amines(e.g. dicyclohexylamine, trimethylamine, triethylamine, pyridine,picoline, ethanolamine, diethanolamine, triethanolamine) or basic aminoacids (e.g. arginine, lysine and omithine). Such salts can be preparedin situ during isolation and purification of the compounds or byseparately reacting the purified compound in its free base or free acidform with a suitable acid or base, respectively, and isolating the saltthus formed.

Those of skill in the art will appreciate that compounds of the presenttechnology may exhibit the phenomena of tautomerism, conformationalisomerism, geometric isomerism and/or stereoisomerism. As the formuladrawings within the specification and claims can represent only one ofthe possible tautomeric, conformational isomeric, stereochemical orgeometric isomeric forms, it should be understood that the presenttechnology encompasses any tautomeric, conformational isomeric,stereochemical and/or geometric isomeric forms of the compounds havingone or more of the utilities described herein, as well as mixtures ofthese various different forms.

“Tautomers” refers to isomeric forms of a compound that are inequilibrium with each other. The presence and concentrations of theisomeric forms will depend on the environment the compound is found inand may be different depending upon, for example, whether the compoundis a solid or is in an organic or aqueous solution. For example, inaqueous solution, quinazolinones may exhibit the following isomericforms, which are referred to as tautomers of each other:

As another example, guanidines may exhibit the following isomeric formsin protic organic solution, also referred to as tautomers of each other:

Because of the limits of representing compounds by structural formulas,it is to be understood that all chemical formulas of the compoundsdescribed herein represent all tautomeric forms of compounds and arewithin the scope of the present technology.

Stereoisomers of compounds (also known as optical isomers) include allchiral, diastereomeric, and racemic forms of a structure, unless thespecific stereochemistry is expressly indicated. Thus, compounds used inthe present technology include enriched or resolved optical isomers atany or all asymmetric atoms as are apparent from the depictions. Bothracemic and diastereomeric mixtures, as well as the individual opticalisomers can be isolated or synthesized so as to be substantially free oftheir enantiomeric or diastereomeric partners, and these stereoisomersare all within the scope of the present technology.

The compounds of the present technology may exist as solvates,especially hydrates. Hydrates may form during manufacture of thecompounds or compositions comprising the compounds, or hydrates may formover time due to the hygroscopic nature of the compounds. Compounds ofthe present technology may exist as organic solvates as well, includingDMF, ether, and alcohol solvates among others. The identification andpreparation of any particular solvate is within the skill of theordinary artisan of synthetic organic or medicinal chemistry.

Activation of the kappa opioid receptor (KOR) by endogenousneuropeptides, primarily dynorphin, initiates complex signalingcascades. The downstream effects of KOR agonism vary greatly and includeantinociception, dysphoria and anxiety, though the details of thepharmacological pathways are still being elucidated. In contrast, KORantagonists have been investigated as therapeutic treatments foraddiction, diuresis, depression, post-traumatic stress disorder, aneating disorder, panic disorder, social anxiety disorder, generalanxiety disorder, obsessive compulsive disorders, excessive orunreasonable specific phobias, as well as other conditions related toanxiety or aversion-reward responses. Many canonical KOR antagonists(Scheme 1) are derived from or bear a structural element of morphinanopioids, such as the widely-utilized tool compounds norBNI, 5′-GNTI andJDTic.

Furthermore, many known KOR antagonists such as norBNI and JDTic arereported to have an extended duration of action, with effects of asingle injection imparting antagonism in rodent models from a period ofdays up to months. There has been some controversy as to what causesthese long term effects, whether it is due to adaptive signalingdownstream of the antagonists (“adaptive plasticity”), or a depot effectwherein the compound is retained in tissues. For example, norBNI hasbeen detected in brain tissue after more than 21 days (Patkar, K. A.;Wu, J.; Ganno, M. L.; Singh, H. D.; Ross, N. C.; Rasakham, K.; Toll, L.;McLaughlin, J. P. Physical presence of nor-binaltorphimine in mousebrain over 21 days after a single administration corresponds to itslong-lasting antagonistic effect on kappa opioid receptors. J.Pharmacol. Exp. Ther. 2013, 346, 545-554.). However, irrespective ofwhether there is a depot effect, adaptive plasticity, or some othermechanism, such extended durations limit the therapeutic uses of suchcompounds in patients.

In contrast to the better-established chemotypes noted, the compounds ofthe present technology are highly modular, possesses no stereogeniccenters, and bear little structural similarity to morphinan-like opioidligands. Moreover, the compounds of the present technology have aclearance rate of less than about 12 h, a duration of action that allowsfor a more safe and effective therapeutic regime for KOR antagonists.

In an aspect, a compound according to formula I is provided

or stereoisomers, tautomers, solvates, and/or salts thereof, where G¹and G² are each independently C═O or S(O)₂; R¹, R², R³, R⁴, R⁵, R⁷, R⁸,R⁹, R¹⁰, and R¹¹ are each independently H, halo, hydroxy, amino, cyano,trifluoromethyl, thiol, alkylthio, sulfoxide, sulfone, nitro,pentafluorosulfanyl, carboxylate, amide, ester, or a substituted orunsubstituted C₁-C₆ alkyl, C₁-C₆ alkoxy, aryl, aryloxy, C₁-C₆ alkanoyl,C₁-C₈ alkanoyloxy, aryloyl, or aryloyloxy group, where any two adjacentR¹, R², R³, R⁴, R⁵, R⁷, R⁸, R⁹, R¹⁰, and R¹¹ may join to form a5-membered or 6-membered substituted or unsubstituted heteroalkyl group;R⁶ is a branched C₁-C₈ alkyl group or a substituted or unsubstitutedcycloalkyl or aryl group; R¹² and R¹³ are each independently H or asubstituted or unsubstituted C₁-C₈ alkyl or C₅-C₇ cycloalkyl group; andn is 0, 1, or 2; provided that when G¹ is S(O)₂, G² is C═O, R¹, R², R⁴,R⁵, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³ are each H, R⁶ is isopropyl, and n is1, then R³ is not methyl.

In any embodiment herein, it may be at least one of R¹, R², R³, R⁴, R⁵,R⁷, R⁸, R⁹, R¹⁰, or R¹¹ is halo, hydroxy, cyano, trifluoromethyl, thiol,alkylthio, nitro, pentafluorosulfanyl, carboxylate, ester, or asubstituted or unsubstituted C₁-C₆ alkyl, C₁-C₆ alkoxy, aryl, aryloxy,C₁-C₆ alkanoyl, C₁-C₆ alkanoyloxy, aryloyl, or aryloyloxy group. In maybe that at least one of R¹, R², R³, R⁴, R⁵, R⁷, R⁸, R⁹, R¹⁰, or R¹¹ ishalo, hydroxy, cyano, trifluoromethyl, thiol, alkylthio, sulfoxide,sulfone, pentafluorosulfanyl, carboxylate, ester, or a substituted orunsubstituted C₁-C₆ alkoxy, aryl, aryloxy, C₁-C₆ alkanoyl, C₁-C₆alkanoyloxy, aryloyl, or aryloyloxy group. In any embodiment herein, itmay be that at least one of R¹, R², R³, R⁴, R⁵, R⁷, R⁸, R⁹, R¹⁰, or R¹¹is halo, hydroxy, cyano, alkylthio, sulfone, carboxylate, ester, or asubstituted or unsubstituted C₁-C₆ alkoxy, aryloxy, C₁-C₆ alkanoyl,C₁-C₆ alkanoyloxy, aryloyl, or aryloyloxy group. In any embodimentherein, it may be that at least one of R¹, R², R³, R⁴, R⁵, R⁷, R⁸, R⁹,R¹⁰, or R¹¹ is halo, hydroxy, carboxylate, ester, or a substituted orunsubstituted C₁-C₆ alkoxy, C₁-C₆ alkanoyl, or C₁-C₆ alkanoyloxy group.In any embodiment herein, it may be that at least one of R¹, R², R³, R⁴,R⁵, R⁷, R⁸, R⁹, R¹⁰, or R¹¹ is hydroxy or a substituted or unsubstitutedC₁-C₆ alkoxy or C₁-C₆ alkanoyloxy group. In any embodiment herein, itmay be that at least one of R¹, R², R³, R⁴, R⁵, R⁷, R⁸, R⁹, R¹⁰, or R¹¹is hydroxy or an unsubstituted C₁-C₆ alkoxy group.

At least one of R¹, R², R³, R⁴, or R⁵ may be halo, hydroxy, carboxylate,ester, or a substituted or unsubstituted C₁-C₆ alkyl, C₁-C₆ alkoxy,C₁-C₆ alkanoyl, or C₁-C₆ alkanoyloxy group. In any embodiment herein, atleast one of R⁷, R⁸, R⁹, R¹⁰, or R¹¹ may be halo, hydroxy, cyano,trifluoromethyl, thiol, alkylthio, sulfoxide, sulfone, nitro,pentafluorosulfanyl, carboxylate, ester, or a substituted orunsubstituted C₁-C₆ alkyl, C₁-C₆ alkoxy, aryl, aryloxy, C₁-C₆ alkanoyl,C₁-C₆ alkanoyloxy, aryloyl, or aryloyloxy group. In any embodimentherein, it may be that at least one of R⁷, R⁸, R⁹, R¹⁰, or R¹¹ is halo,hydroxy, trifluoromethyl, carboxylate, ester, or a substituted orunsubstituted C₁-C₆ alkyl, C₁-C₆ alkoxy, C₁-C₆ alkanoyl, or C₁-C₆alkanoyloxy group. In any embodiment herein, it may be that at least oneof R⁷, R⁸, R⁹, R¹⁰, or R¹¹ is hydroxy or an unsubstituted C₁-C₆ alkoxygroup.

At least two of R¹, R², R³, R⁴, R⁵, R⁷, R⁸, R⁹, R¹⁰, or R¹¹ may eachindependently be halo, hydroxy, cyano, trifluoromethyl, nitro,pentafluorosulfanyl, carboxylate, ester, or a substituted orunsubstituted C₁-C₆ alkyl, C₁-C₆ alkoxy, aryl, aryloxy, C₁-C₆ alkanoyl,C₁-C₆ alkanoyloxy, aryloyl, or aryloyloxy group. It may be that at leasttwo of R¹, R², R³, R⁴, R⁵, R⁷, R⁸, R⁹, R¹⁰, or R¹¹ are eachindependently halo, hydroxy, carboxylate, ester, or a substituted orunsubstituted C₁-C₆ alkyl, C₁-C₆ alkoxy, C₁-C₆ alkanoyl, or C₁-C₆alkanoyloxy group. In any embodiment herein, it may be that at least twoof R¹, R², R³, R⁴, R⁵, R⁷, R⁸, R⁹, R¹⁰, or R¹¹ are each independentlyhalo, hydroxy, or a substituted or unsubstituted C₁-C₆ alkyl or C₁-C₆alkoxy group.

In any of the above embodiments, it may be that one of R¹, R², R³, R⁴ orR⁵ is halo, hydroxy, amino, cyano, trifluoromethyl, nitro,pentafluorosulfanyl, carboxylate, amide, ester, or a substituted orunsubstituted C₁-C₆ alkyl, C₁-C₆ alkoxy, aryl, aryloxy, C₁-C₆ alkanoyl,C₁-C₆ alkanoyloxy, aryloyl, or aryloyloxy group; and the remaining R¹,R², R³, R⁴, or R⁵ are each H. In any of the above embodiments, it may bethat any two adjacent R¹, R², R³, R⁴, R⁵, R⁷, R⁸, R⁹, R¹⁰, and R¹¹ mayjoin to form a dioxolanyl or dioxanyl group. In any of the aboveembodiments, it may be that n is 1. In any of the above embodiments, itmay be that G¹ is S(O)₂. In any of the above embodiments, it may be thatG² is C═O. In any of the above embodiments, it may be that R⁶ is abranched C₁-C₈ alkyl group or a substituted or unsubstituted cycloalkylgroup. In any of the above embodiments, it may be that R⁶ is isopropyl,sec-butyl, tert-butyl, isopentyl, neopentyl, or adamantyl. In any of theabove embodiments, it may be that R⁶ is tert-butyl, neopentyl, oradamantyl.

In any of the above embodiments, it may be that when G¹ is S(O)₂, G² isC═O, R¹, R², R⁴, R⁵, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³ are each H, and R⁶is isopropyl, then R³ is not an unsubstituted linear alkyl group. In anyof the above embodiments, it may be that when G¹ is S(O)₂, G² is C═O,R¹, R², R⁴, R⁵, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³ are each H, and R⁶ isisopropyl, then R³ is not an unsubstituted alkyl group. In any of theabove embodiments, it may be that when G¹ is S(O)₂, G² is C═O, R¹, R²,R⁴, R⁵, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³ are each H, and R⁶ is isopropyl,then R³ is not an alkyl group.

In any embodiment herein, it may be that G¹ is S(O)₂; G² is C═O; R¹, R⁴,R⁵, R⁷, R¹⁰, and R¹¹ are each H; R², R³, R⁸, and R⁹ are eachindependently halo, hydroxy, amino, cyano, trifluoromethyl, thiol,alkylthio, sulfoxide, sulfone, nitro, pentafluorosulfanyl, carboxylate,amide, ester, or a substituted or unsubstituted C₁-C₆ alkyl, C₁-C₆alkoxy, aryl, aryloxy, C₁-C₆ alkanoyl, C₁-C₈ alkanoyloxy, aryloyl, oraryloyloxy group, where any two adjacent R², R³, R⁸, and R⁹ may join toform a 5-membered or 6-membered substituted or unsubstituted heteroalkylgroup; R⁶ is a branched C₁-C₈ alkyl group; R¹² and R¹³ are each H; and nis 1.

In any embodiment herein, it may be that G¹ is S(O)₂; G² is C═O; R¹, R⁴,R⁵, R⁷, R¹⁰, and R¹¹ are each H; one of R² and R³ is halo, hydroxy, or aunsubstituted C₁-C₆ alkyl, C₁-C₆ alkoxy, C₁-C₆ alkanoyl, or C₁-C₈alkanoyloxy group and the other R² or R³ is H; one of R⁸ and R⁹ is halo,hydroxy, amino, cyano, trifluoromethyl, thiol, alkylthio, sulfoxide,sulfone, nitro, pentafluorosulfanyl, carboxylate, amide, ester, or asubstituted or unsubstituted C₁-C₆ alkoxy, C₁-C₆ alkanoyl, or C₁-C₈alkanoyloxy group, and the other R⁸ or R⁹ is H; R⁶ is a branched C₁-C₈alkyl group; R¹² and R¹³ are each H; and n is 1.

In any of the above embodiments, it may be that the compound is selectedfrom

In any of the above embodiments, it may be that the compound is selectedfrom

In an aspect of the present technology, a composition is provided thatincludes any one of the aspects and embodiments of compounds of formulaI and a pharmaceutically acceptable carrier. In a related aspect, apharmaceutical composition is provided, the pharmaceutical compositionincluding an effective amount of the compound of any one of the aspectsand embodiments of compounds of formula I for treating a condition; andwhere the condition is addiction, diuresis, depression, post-traumaticstress disorder, an eating disorder, panic disorder, social anxietydisorder, general anxiety disorder, obsessive compulsive disorders,excessive or unreasonable specific phobias, and/or other conditionsrelated to anxiety or aversion-reward responses. In a further relatedaspect, a method is provided that includes administering an effectiveamount of a compound of any one of the aspects and embodiments ofcompounds of formula I or administering a pharmaceutical compositioncomprising an effective amount of a compound of any one of the aspectsand embodiments of compounds of formulas I to a subject suffering fromaddiction, diuresis, depression, post traumatic stress disorder, aneating disorder, panic disorder, social anxiety disorder, generalanxiety disorder, obsessive compulsive disorders, excessive orunreasonable specific phobias, and/or other conditions related toanxiety or aversion-reward responses.

“Effective amount” refers to the amount of a compound or compositionrequired to produce a desired effect. One example of an effective amountincludes amounts or dosages that yield acceptable toxicity andbioavailability levels for therapeutic (pharmaceutical) use including,but not limited to, the treatment of alcohol addiction. Another exampleof an effective amount includes amounts or dosages that are capable ofreducing symptoms associated with metabolic syndrome, such as, forexample, obesity and/or cardiometabolic abnormalities. The effectiveamount of the compound may selectively antagonize the kappa opioidreceptor (KOR). The effective amount of the compound may selectivelybind to the KOR at least about 5 times more than the 6 opioid receptor(DOR); thus, the effective amount of the compound may selectively bindto the KOR at least about 10 times, at least about 25 times, at leastabout 50 times, or at least about 100 times more than the DOR. In anyembodiment herein, including any of the above embodiments regarding theDOR, the effective amount of the compound may selectively bind to theKOR at least about 5 times more than the g opioid receptor (MOR); thus,it may be that the effective amount of the compound selectively binds tothe KOR at least about 10 times more, at least 25 times more, at leastabout 50 times more, or at least about 100 times more than the MOR. Asused herein, a “subject” or “patient” is a mammal, such as a cat, dog,rodent or primate. Typically the subject is a human, and, preferably, ahuman suffering from or suspected of suffering from an addiction. Theterm “subject” and “patient” can be used interchangeably.

Thus, the instant present technology provides pharmaceuticalcompositions and medicaments comprising any of the compounds disclosedherein (e.g., compounds of formulas I) and a pharmaceutically acceptablecarrier or one or more excipients or fillers. The compositions may beused in the methods and treatments described herein. Such compositionsand medicaments include a therapeutically effective amount of anycompound as described herein, including but not limited to a compound offormula I. The pharmaceutical composition may be packaged in unit dosageform. The unit dosage form is effective in treating addiction byreducing desire for an addictive substance(s), and/or effective intreating a metabolic disorder by reducing symptoms associated with themetabolic disorder when administered to a subject in need thereof.

The pharmaceutical compositions and medicaments may be prepared bymixing one or more compounds of the present technology, pharmaceuticallyacceptable salts thereof, stereoisomers thereof, tautomers thereof, orsolvates thereof, with pharmaceutically acceptable carriers, excipients,binders, diluents or the like to prevent and treat disorders associatedwith the effects of increased plasma and/or hepatic lipid levels. Thecompounds and compositions described herein may be used to prepareformulations and medicaments that prevent or treat a variety ofdisorders associated with addiction, diuresis, depression,post-traumatic stress disorder, an eating disorder, panic disorder,social anxiety disorder, general anxiety disorder, obsessive compulsivedisorders, excessive or unreasonable specific phobias, and/or otherconditions related to anxiety or aversion-reward responses. Suchcompositions can be in the form of, for example, granules, powders,tablets, capsules, syrup, suppositories, injections, emulsions, elixirs,suspensions or solutions. The instant compositions can be formulated forvarious routes of administration, for example, by oral, parenteral,topical, rectal, nasal, vaginal administration, or via implantedreservoir. Parenteral or systemic administration includes, but is notlimited to, subcutaneous, intravenous, intraperitoneal, andintramuscular, injections. The following dosage forms are given by wayof example and should not be construed as limiting the instant presenttechnology.

For oral, buccal, and sublingual administration, powders, suspensions,granules, tablets, pills, capsules, gelcaps, and caplets are acceptableas solid dosage forms. These can be prepared, for example, by mixing oneor more compounds of the instant present technology, or pharmaceuticallyacceptable salts or tautomers thereof, with at least one additive suchas a starch or other additive. Suitable additives are sucrose, lactose,cellulose sugar, mannitol, maltitol, dextran, starch, agar, alginates,chitins, chitosans, pectins, tragacanth gum, gum arabic, gelatins,collagens, casein, albumin, synthetic or semi-synthetic polymers orglycerides. Optionally, oral dosage forms can contain other ingredientsto aid in administration, such as an inactive diluent, or lubricantssuch as magnesium stearate, or preservatives such as paraben or sorbicacid, or anti-oxidants such as ascorbic acid, tocopherol or cysteine, adisintegrating agent, binders, thickeners, buffers, sweeteners,flavoring agents or perfuming agents. Tablets and pills may be furthertreated with suitable coating materials known in the art.

Liquid dosage forms for oral administration may be in the form ofpharmaceutically acceptable emulsions, syrups, elixirs, suspensions, andsolutions, which may contain an inactive diluent, such as water.Pharmaceutical formulations and medicaments may be prepared as liquidsuspensions or solutions using a sterile liquid, such as, but notlimited to, an oil, water, an alcohol, and combinations of these.Pharmaceutically suitable surfactants, suspending agents, emulsifyingagents, may be added for oral or parenteral administration.

As noted above, suspensions may include oils. Such oils include, but arenot limited to, peanut oil, sesame oil, cottonseed oil, corn oil andolive oil. Suspension preparation may also contain esters of fatty acidssuch as ethyl oleate, isopropyl myristate, fatty acid glycerides andacetylated fatty acid glycerides. Suspension formulations may includealcohols, such as, but not limited to, ethanol, isopropyl alcohol,hexadecyl alcohol, glycerol and propylene glycol. Ethers, such as butnot limited to, poly(ethyleneglycol), petroleum hydrocarbons such asmineral oil and petrolatum; and water may also be used in suspensionformulations.

Injectable dosage forms generally include aqueous suspensions or oilsuspensions which may be prepared using a suitable dispersant or wettingagent and a suspending agent. Injectable forms may be in solution phaseor in the form of a suspension, which is prepared with a solvent ordiluent. Acceptable solvents or vehicles include sterilized water,Ringer's solution, or an isotonic aqueous saline solution.Alternatively, sterile oils may be employed as solvents or suspendingagents. Typically, the oil or fatty acid is non-volatile, includingnatural or synthetic oils, fatty acids, mono-, di- or tri-glycerides.

For injection, the pharmaceutical formulation and/or medicament may be apowder suitable for reconstitution with an appropriate solution asdescribed above. Examples of these include, but are not limited to,freeze dried, rotary dried or spray dried powders, amorphous powders,granules, precipitates, or particulates. For injection, the formulationsmay optionally contain stabilizers, pH modifiers, surfactants,bioavailability modifiers and combinations of these.

Compounds of the present technology may be administered to the lungs byinhalation through the nose or mouth. Suitable pharmaceuticalformulations for inhalation include solutions, sprays, dry powders, oraerosols containing any appropriate solvents and optionally othercompounds such as, but not limited to, stabilizers, antimicrobialagents, antioxidants, pH modifiers, surfactants, bioavailabilitymodifiers and combinations of these. The carriers and stabilizers varywith the requirements of the particular compound, but typically includenonionic surfactants (Tweens, Pluronics, or polyethylene glycol),innocuous proteins like serum albumin, sorbitan esters, oleic acid,lecithin, amino acids such as glycine, buffers, salts, sugars or sugaralcohols. Aqueous and nonaqueous (e.g., in a fluorocarbon propellant)aerosols are typically used for delivery of compounds of the presenttechnology by inhalation.

Dosage forms for the topical (including buccal and sublingual) ortransdermal administration of compounds of the present technologyinclude powders, sprays, ointments, pastes, creams, lotions, gels,solutions, and patches. The active component may be mixed under sterileconditions with a pharmaceutically-acceptable carrier or excipient, andwith any preservatives, or buffers, which may be required. Powders andsprays can be prepared, for example, with excipients such as lactose,talc, silicic acid, aluminum hydroxide, calcium silicates and polyamidepowder, or mixtures of these substances. The ointments, pastes, creamsand gels may also contain excipients such as animal and vegetable fats,oils, waxes, paraffins, starch, tragacanth, cellulose derivatives,polyethylene glycols, silicones, bentonites, silicic acid, talc and zincoxide, or mixtures thereof. Absorption enhancers can also be used toincrease the flux of the compounds of the present technology across theskin. The rate of such flux can be controlled by either providing a ratecontrolling membrane (e.g., as part of a transdermal patch) ordispersing the compound in a polymer matrix or gel.

Besides those representative dosage forms described above,pharmaceutically acceptable excipients and carriers are generally knownto those skilled in the art and are thus included in the instant presenttechnology. Such excipients and carriers are described, for example, in“Remingtons Pharmaceutical Sciences” Mack Pub. Co., New Jersey (1991),which is incorporated herein by reference.

The formulations of the present technology may be designed to beshort-acting, fast-releasing, long-acting, and sustained-releasing asdescribed below. Thus, the pharmaceutical formulations may also beformulated for controlled release or for slow release.

The instant compositions may also comprise, for example, micelles orliposomes, or some other encapsulated form, or may be administered in anextended release form to provide a prolonged storage and/or deliveryeffect. Therefore, the pharmaceutical formulations and medicaments maybe compressed into pellets or cylinders and implanted intramuscularly orsubcutaneously as depot injections or as implants such as stents. Suchimplants may employ known inert materials such as silicones andbiodegradable polymers.

Specific dosages may be adjusted depending on conditions of disease, theage, body weight, general health conditions, sex, and diet of thesubject, dose intervals, administration routes, excretion rate, andcombinations of drugs. Any of the above dosage forms containingeffective amounts are well within the bounds of routine experimentationand therefore, well within the scope of the instant present technology.

Those skilled in the art are readily able to determine an effectiveamount by simply administering a compound of the present technology to apatient in increasing amounts until (for addiction) the motivation tointernalize the addictive substance and/or relapse-like behavior isdecreased or stopped, or (for metabolic syndrome and/or obesity) theelevated plasma or elevated white blood cell count or hepaticcholesterol or triglycerides or progression of the disease state isdecreased or stopped. For metabolic syndrome and/or obesity, theprogression of the disease state can be assessed using in vivo imaging,as described, or by taking a tissue sample from a patient and observingthe target of interest therein. The compounds of the present technologycan be administered to a patient at dosage levels in the range of about0.1 to about 1,000 mg per day. For a normal human adult having a bodyweight of about 70 kg, a dosage in the range of about 0.01 to about 100mg per kg of body weight per day is sufficient. The specific dosageused, however, can vary or may be adjusted as considered appropriate bythose of ordinary skill in the art. For example, the dosage can dependon a number of factors including the requirements of the patient, theseverity of the condition being treated and the pharmacological activityof the compound being used. The determination of optimum dosages for aparticular patient is well known to those skilled in the art.

Various assays and model systems can be readily employed to determinethe therapeutic effectiveness of the treatment according to the presenttechnology.

Effectiveness of the compositions and methods of the present technologymay also be demonstrated by a decrease in the symptoms of an addiction,such as, for example, motivation to internalize the addictive substanceand/or relapse-like behavior. Effectiveness of the compositions andmethods of the present technology may also be demonstrated by a decreasein the symptoms of addiction, diuresis, depression, post-traumaticstress disorder, an eating disorder, panic disorder, social anxietydisorder, general anxiety disorder, obsessive compulsive disorders,excessive or unreasonable specific phobias, and/or other conditionsrelated to anxiety or aversion-reward responses.

For each of the indicated conditions described herein, test subjectswill exhibit a 10%, 20%, 30%, 50% or greater reduction, up to a 75-90%,or 95% or greater, reduction, in one or more symptom(s) caused by, orassociated with, the disorder in the subject, compared toplacebo-treated or other suitable control subjects.

In an aspect, a method is provided where the method includes inhibitingβ-arrestin recruitment in a subject by administering an effective amountof a compound of any one of the aspects and embodiments of compounds offormula I. The subject may be suffering from addiction, diuresis,depression, post-traumatic stress disorder, an eating disorder, panicdisorder, social anxiety disorder, general anxiety disorder, obsessivecompulsive disorders, excessive or unreasonable specific phobias, and/orother conditions related to anxiety or aversion-reward responses. In anyof the above embodiments, the addiction may be to at least one ofnicotine, ethanol, cocaine, opioids, amphetamines, marijuana, or asynthetic cannabinoid agonist. In any embodiment herein, the method mayinclude inhibiting β-arrestin2 recruitment.

In an aspect, a method for treating an addiction in a subject isprovided that includes administering an effective amount of a compoundof any one of the aspects and embodiments of compounds of formula I. Theaddiction may be to at least one of nicotine, ethanol, cocaine, opioids,amphetamines, marijuana, or a synthetic cannabinoid agonist.Administering the effective amount of the compound may includeadministering a pharmaceutical composition according to any embodimentdescribed herein.

In an aspect, a method of inhibiting β-arrestin recruitment is providedthat includes contacting a KOR with a compound of any one of the aspectsand embodiments of compounds of formulas I. It may be the methodincludes contacting a KOR with an effective amount of a compound of anyone of the aspects and embodiments of compounds of formulas I. Suchmethods may be performed outside of a subject, such as in an assay. Acell may include the KOR.

The compounds of the present technology can also be administered to apatient along with other conventional therapeutic agents that may beuseful in the treatment of addiction, diuresis, depression,post-traumatic stress disorder, an eating disorder, panic disorder,social anxiety disorder, general anxiety disorder, obsessive compulsivedisorders, excessive or unreasonable specific phobias, and/or otherconditions related to anxiety or aversion-reward responses. Theadministration may include oral administration, parenteraladministration, or nasal administration. In any of these embodiments,the administration may include subcutaneous injections, intravenousinjections, intraperitoneal injections, or intramuscular injections. Inany of these embodiments, the administration may include oraladministration. The methods of the present technology can also compriseadministering, either sequentially or in combination with one or morecompounds of the present technology, a conventional therapeutic agent inan amount that can potentially or synergistically be effective for thetreatment of addiction, diuresis, depression, post traumatic stressdisorder, an eating disorder, panic disorder, social anxiety disorder,general anxiety disorder, obsessive compulsive disorders, excessive orunreasonable specific phobias, and/or other conditions related toanxiety or aversion-reward responses.

In one aspect, a compound of the present technology is administered to apatient in an amount or dosage suitable for therapeutic use. Generally,a unit dosage comprising a compound of the present technology will varydepending on patient considerations. Such considerations include, forexample, age, protocol, condition, sex, extent of disease,contraindications, concomitant therapies and the like. An exemplary unitdosage based on these considerations can also be adjusted or modified bya physician skilled in the art. For example, a unit dosage for a patientcomprising a compound of the present technology can vary from 1×10⁻⁴g/kg to 1 g/kg, preferably, 1×10⁻³ g/kg to 1.0 g/kg. Dosage of acompound of the present technology can also vary from 0.01 mg/kg to 100mg/kg or, preferably, from 0.1 mg/kg to 10 mg/kg.

A compound of the present technology can also be modified, for example,by the covalent attachment of an organic moiety or conjugate to improvepharmacokinetic properties, toxicity or bioavailability (e.g., increasedin vivo half-life). The conjugate can be a linear or branchedhydrophilic polymeric group, fatty acid group or fatty acid ester group.A polymeric group can comprise a molecular weight that can be adjustedby one of ordinary skill in the art to improve, for example,pharmacokinetic properties, toxicity or bioavailability. Exemplaryconjugates can include a polyalkane glycol (e.g., polyethylene glycol(PEG), polypropylene glycol (PPG)), carbohydrate polymer, amino acidpolymer or polyvinyl pyrolidone and a fatty acid or fatty acid estergroup, each of which can independently comprise from about eight toabout seventy carbon atoms. Conjugates for use with a compound of thepresent technology can also serve as linkers to, for example, anysuitable substituents or groups, radiolabels (marker or tags), halogens,proteins, enzymes, polypeptides, other therapeutic agents (for example,a pharmaceutical or drug), nucleosides, dyes, oligonucleotides, lipids,phospholipids and/or liposomes. In one aspect, conjugates can includepolyethylene amine (PEI), polyglycine, hybrids of PEI and polyglycine,polyethylene glycol (PEG) or methoxypolyethylene glycol (mPEG). Aconjugate can also link a compound of the present technology to, forexample, a label (fluorescent or luminescent) or marker (radionuclide,radioisotope and/or isotope) to comprise a probe of the presenttechnology. Conjugates for use with a compound of the present technologycan, in one aspect, improve in vivo half-life. Other exemplaryconjugates for use with a compound of the present technology as well asapplications thereof and related techniques include those generallydescribed by U.S. Pat. No. 5,672,662, which is hereby incorporated byreference herein.

In another aspect, the present technology provides methods ofidentifying a target of interest including contacting the target ofinterest with a detectable or imaging effective quantity of a labeledcompound of the present technology. A detectable or imaging effectivequantity is a quantity of a labeled compound of the present technologynecessary to be detected by the detection method chosen. For example, adetectable quantity can be an administered amount sufficient to enabledetection of binding of the labeled compound to a target of interestincluding, but not limited to, a KOR. Suitable labels are known by thoseskilled in the art and can include, for example, radioisotopes,radionuclides, isotopes, fluorescent groups, biotin (in conjunction withstreptavidin complexation), and chemiluminescent groups. Upon binding ofthe labeled compound to the target of interest, the target may beisolated, purified and further characterized such as by determining theamino acid sequence.

The terms “associated” and/or “binding” can mean a chemical or physicalinteraction, for example, between a compound of the present technologyand a target of interest. Examples of associations or interactionsinclude covalent bonds, ionic bonds, hydrophilic-hydrophilicinteractions, hydrophobic-hydrophobic interactions and complexes.Associated can also refer generally to “binding” or “affinity” as eachcan be used to describe various chemical or physical interactions.Measuring binding or affinity is also routine to those skilled in theart. For example, compounds of the present technology can bind to orinteract with a target of interest or precursors, portions, fragmentsand peptides thereof and/or their deposits.

The examples herein are provided to illustrate advantages of the presenttechnology and to further assist a person of ordinary skill in the artwith preparing or using the compounds of the present technology orsalts, pharmaceutical compositions, derivatives, solvates, metabolites,prodrugs, racemic mixtures or tautomeric forms thereof. The examplesherein are also presented in order to more fully illustrate thepreferred aspects of the present technology. The examples should in noway be construed as limiting the scope of the present technology, asdefined by the appended claims. The examples can include or incorporateany of the variations, aspects or aspects of the present technologydescribed above. The variations, aspects or aspects described above mayalso further each include or incorporate the variations of any or allother variations, aspects or aspects of the present technology.

EXAMPLES

General synthetic and analytical details:

All reagents and materials were purchased from commercial vendors(Sigma, Alfa Aesar, Oakwood or ASW Medchem) and used as received. Ethylether, toluene, THF, MeCN and CH₂Cl₂ were degassed with nitrogen andpassed through two columns of basic alumina on an Innovative Technologysolvent purification system. ¹H and ¹³C NMR spectra were recorded on aBruker AM 400 spectrometer (operating at 400 and 100 MHz respectively)in CDCl₃ with 0.03% TMS as an internal standard, unless otherwisespecified. Chemical shifts are reported in parts per million (ppm)downfield from TMS. ¹³C multiplicities were determined with the aid ofan APT pulse sequence, differentiating the signals for methyl andmethane carbons as “d” from methylene and quarternary carbons as “u”.The infrared (IR) spectra were acquired as thin films using a universalATR sampling accessory on a PerkinElmer Spectrum 100 FT-IR spectrometerand the absorption frequencies are reported in cm⁻¹. Melting points weredetermined on a Stanford Research Systems Optimelt automated meltingpoint system interfaced through a PC and are uncorrected.

HPLC/MS analysis was carried out with gradient elution (5% CH₃CN to 100%CH₃CN) on an Agilent 1200 RRLC with a photodiode array UV detector andan Agilent 6224 TOF mass spectrometer (also used to produce highresolution mass spectra). HPLC purification was carried out by massdirected fractionation (MDF) with gradient elution (a narrow CH₃CNgradient was chosen based on the retention time of the target from LCMSanalysis of the crude sample) on an Agilent 1200 instrument withphotodiode array detector, an Agilent 6120 quadrupole mass spectrometer,and a HTPAL LEAP autosampler. Fractions were triggered using an MS andUV threshold determined by HPLC/MS analysis of the crude sample. One oftwo column/mobile phase conditions were chosen for both analysis andpurification to promote the targets neutral state (0.02% formic acidwith Waters Atlantis T3 5 um, 19×150 mm; or pH 9.8 NH₄OH with WatersXBridge C18 5 um, 19×150 mm).

Representative General Synthetic Scheme

Synthesis of Carboxylic Acid Fragments 2a-2c

Example 1: Methyl 2-tosyl-1,2,3,4-tetrahydroisoquinoline-6-carboxylate

To a solution of methyl 1,2,3,4-tetrahydroisoquinoline-6-carboxylate(171 mg, 0.894 mmol) and triethylamine (271 mg, 2.68 mmol, 3.0 equiv) inCH₂Cl₂ (15 mL) was added p-toluenesulfonyl chloride (256 mg, 1.34 mmol,1.5 equiv). The reaction was stirred at rt for 15 h, diluted with 1 NHCl and extracted with CH₂Cl₂. The combined organics were dried(Na₂SO₄), concentrated and purified by silica gel chromatography toafford the sulfonamide product as a white solid (231 mg, 0.669 mmol, 75%yield). Mp=143-145° C.; R_(f)=0.34 (25% EtOAc/hexanes); ¹H NMR (400 MHz,CDCl₃) δ 2.41 (s, 3H), 2.96 (t, J=6.0 Hz, 2H), 3.36 (t, J=6.0 Hz, 2H),3.88 (s, 3H), 4.28 (s, 2H), 7.10 (d, J=8.0 Hz, 1H), 7.33 (d, J=8.0 Hz,2H), 7.72 (d, J=8.0 Hz, 2H), 7.77 (s, 1H), 7.79 (d, J=8.0 Hz, 1H); ¹³CNMR (101 MHz, CDCl₃, APT pulse sequence) δ d 21.5, 52.1, 126.6, 127.4,127.7, 129.8, 130.2; u 28.8, 43.6, 47.7, 128.7, 133.2, 133.4, 136.9,143.9, 166.7; IR 1718 cm⁻¹; HRMS (ESI) m/z calcd for C₁₈H₂₀NO₄S ([M+H]⁺)346.1108, found 346.1116.

Example 2: 2-Tosyl-1,2,3,4-tetrahydroisoquinoline-6-carboxylic Acid (2a)

To a solution of the above methyl ester (298 mg, 0.765 mmol) inTHF:MeOH:water (3:1:1, 10 mL) was added lithium hydroxide monohydrate(160 mg, 3.83 mmol, 5 equiv) and the reaction stirred at rt for 15 h.The THF and MeOH were removed in vacuo and the reaction concentrate wasacidified with 2 N HCl, precipitating the carboxylic acid product as asparingly soluble white solid (213 mg, 0.643 mmol, 84% yield), which wasfiltered, washed with water, dried under vacuum and used without furtherpurification. Mp=234-240° C.; R_(f)=0.58 (10% MeOH and 2% AcOH inCH₂Cl₂); ¹H NMR (400 MHz, DMSO-d6) δ 2.39 (s, 3H), 2.91 (t, J=6.0 Hz,2H), 3.30 (t, J=6.0 Hz, 2H), 4.25 (s, 2H), 7.28 (d, J=8.0 Hz, 1H), 7.44(d, J=8.0 Hz, 2H), 7.70-7.74 (complex, 4H), 12.89 (br s, 1H); ¹³C NMR(101 MHz, DMSO-d6, APT pulse sequence) δ d 21.0, 126.7, 126.9, 127.4,129.7, 129.9; u 27.9, 43.3, 47.3, 129.1, 133.0, 133.4, 136.7, 143.7,167.0; IR 1678 cm⁻¹; HRMS (ESI) m/z calcd for C₁₇H₁₆NO₄S ([M−H]⁻)330.0806, found 330.0807.

Example 3: Methyl2-((4-ethylphenyl)sulfonyl)-1,2,3,4-tetrahydroisoquinoline-6-carboxylate

Methyl 1,2,3,4-tetrahydroisoquinoline-6-carboxylate (182 mg, 0.952 mmol)and 4-ethyl-benzenesulfonyl chloride (292 mg, 1.43 mmol, 1.5 equiv) werereacted according the protocol in Example 1 to afford the sulfonamideproduct as a white solid (285 mg, 0.793 mmol, 83% yield). Mp=128-130°C.; R_(f)=0.40 (25% EtOAc/hexanes); ¹H NMR (400 MHz, CDCl₃) δ 1.23 (t,J=7.6 Hz, 3H), 2.69 (q, J=7.6 Hz, 2H), 2.95 (t, J=6.0 Hz, 2H), 3.36 (t,J=6.0 Hz, 2H), 3.87 (s, 3H), 4.28 (s, 2H), 7.09 (d, J=8.4 Hz, 1H), 7.35(d, J=8.4 Hz, 2H), 7.74-7.79 (complex, 4H); ¹³C NMR (101 MHz, CDCl₃, APTpulse sequence) δ d 14.9, 51.9, 126.4, 127.2, 127.7, 128.5, 130.0; u28.62, 28.65, 43.5, 47.6, 128.6, 133.2, 133.3, 136.8, 149.8, 166.5; IR1717 cm⁻¹; HRMS (ESI) m/z calcd for C₁₉H₂₂NO₄S ([M+H]⁺) 360.1264, found360.1274.

Example 4:2-((4-Ethylphenyl)sulfonyl)-1,2,3,4-tetrahydroisoquinoline-6-carboxylicAcid (2b)

The above methyl ester (252 mg, 0.701 mmol) were reacted according theprotocol in Example 2 to afford the carboxylic acid product as asparingly soluble white solid (216 mg, 0.625 mmol, 89% yield), which wasfiltered, washed with water, dried under vacuum and used without furtherpurification. Mp=208-213° C.; R_(f)=0.58 (10% MeOH and 2% AcOH inCH₂Cl₂); ¹H NMR (400 MHz, DMSO-d6) δ 1.18 (t, J=7.6 Hz, 3H), 2.68 (q,J=7.6 Hz, 2H), 2.91 (t, J=6.0 Hz, 2H), 3.31 (t, J=6.0 Hz, 2H), 4.26 (s,2H), 7.28 (d, J=8.0 Hz, 1H), 7.46 (d, J=8.0 Hz, 2H), 7.69-7.76 (complex,4H), 12.91 (br s, 1H); ¹³C NMR (101 MHz, DMSO-d6, APT pulse sequence) δd 15.0, 126.7, 126.8, 127.5, 128.7, 129.7; u 28.0, 43.3, 47.3, 129.1,133.2, 133.4, 136.6, 149.6, 167.0; IR 1683 cm⁻¹; HRMS (ESI) m/z calcdfor C₁₈H₂₀NO₄S ([M+H]⁺) 346.1113, found 346.1114.

Example 5: Methyl2-((2,4,6-trimethylphenyl)sulfonyl)-1,2,3,4-tetrahydroisoquinoline-6-carboxylate

Methyl 1,2,3,4-tetrahydroisoquinoline-6-carboxylate (191 mg, 0.999 mmol)and 2,4,6-trimethyl-benzenesulfonyl chloride (328 mg, 1.50 mmol, 1.5equiv) were reacted according the protocol in Example 1 to afford thesulfonamide product as a white solid (188 mg, 0.503 mmol, 50% yield).Mp=142-143° C.; R_(f)=0.50 (25% EtOAc/hexanes); ¹H NMR (400 MHz, CDCl₃)δ 2.29 (s, 3H), 2.63 (s, 6H), 2.91 (t, J=6.0 Hz, 2H), 3.47 (t, J=6.0 Hz,2H), 3.89 (s, 3H), 4.40 (s, 2H), 6.96 (s, 2H), 7.12 (d, J=8.0 Hz, 1H),7.81 (m, 2H); ¹³C NMR (101 MHz, CDCl₃, APT pulse sequence) δ d 21.0,22.8, 52.1, 126.6, 127.4, 130.3, 132.0; u 28.6, 41.9, 45.9, 128.7,131.7, 133.8, 137.3, 140.5, 142.9, 166.7; IR 1718 cm⁻¹; HRMS (ESI) m/zcalcd for C₂₀H₂₄NO₄S ([M+H]⁺) 374.1426, found 374.1424.

Example 6:2-((2,4,6-Trimethylphenyl)sulfonyl)-1,2,3,4-tetrahydroisoquinoline-6-carboxylicAcid (2c)

The above methyl ester (146 mg, 0.391 mmol) were reacted according theprotocol in Example 2 to afford the carboxylic acid product as asparingly soluble white solid (137 mg, 0.381 mmol, 97% yield), which wasfiltered, washed with water, dried under vacuum and used without furtherpurification. Mp=222-233° C.; R_(f)=0.55 (10% MeOH and 2% AcOH inCH₂Cl₂); ¹H NMR (400 MHz, DMSO-d6) δ 2.28 (s, 3H), 2.55 (s, 6H), 2.87(t, J=5.6 Hz, 2H), 3.43 (t, J=5.6 Hz, 2H), 4.36 (s, 2H), 7.08 (s, 2H),7.31 (d, J=7.6 Hz, 1H), 7.71-7.73 (m, 2H); ¹³C NMR (101 MHz, DMSO-d6,APT pulse sequence) δ d 20.5, 22.3, 126.8, 126.9, 129.9, 131.9; u 27.8,41.6, 45.5, 129.2, 133.8, 137.2, 139.6, 142.6, 167.1; IR 1683 cm⁻¹; HRMS(ESI) m/z calcd for C₁₉H₂₂NO₄S ([M+H]⁺) 360.1264, found 360.1270.

Conversion of Carboxylic Acids 2a-2c to the Acid Chlorides

Example 7: 2-Tosyl-1,2,3,4-tetrahydroisoquinoline-6-carbonyl chloride

The carboxylic acid 2a (145 mg, 0.438 mmol) was dissolved in thionylchloride (0.95 mL, 13.13 mmol, 30 equiv) and heated at 65° C. for 4 h.Excess thionyl chloride was removed in vacuo and the residueazeotropically dried with toluene (3×5 mL) to afford the acid chlorideas a white solid (147 mg, 0.420 mmol, 96% yield), which was used withoutfurther purification. ¹H NMR (400 MHz, CDCl₃) δ 2.36 (s, 3H), 2.94 (t,J=5.9 Hz, 2H), 3.32 (t, J=5.9 Hz, 2H), 4.25 (s, 2H), 7.11 (d, J=8.2 Hz,1H), 7.25-7.30 (m, 2H), 7.63-7.69 (m, 2H), 7.77-7.85 (m, 2H).

Example 8:2-((4-Ethylphenyl)sulfonyl)-1,2,3,4-tetrahydroisoquinoline-6-carbonylchloride

The carboxylic acid 2b (303 mg, 0.878 mmol) was reacted according theprotocol in Example 7 to afford the acid chloride as a white solid (322mg, 0.860 mmol, 98% yield), which was used without further purification.¹H NMR (400 MHz, CDCl₃) δ 1.19 (t, J=7.6 Hz, 3H), 2.65 (q, J=7.6 Hz,2H), 2.95 (t, J=5.9 Hz, 2H), 3.33 (t, J=5.9 Hz, 2H), 4.25 (s, 2H), 7.11(d, J=8.2 Hz, 1H), 7.27-7.32 (m, 2H), 7.65-7.71 (m, 2H), 7.77-7.85 (m,2H).

Example 9: 2-(Mesitylsulfonyl)-1,2,3,4-tetrahydroisoquinoline-6-carbonylchloride

The carboxylic acid 2c (38 mg, 0.106 mmol) was reacted according theprotocol in Example 7 to afford the acid chloride as a white solid (39mg, 0.103 mmol, 98% yield), which was used without further purification.¹H NMR (400 MHz, CDCl₃) δ 2.31 (s, 3H), 2.63 (s, 6H), 2.95 (t, J=6.0 Hz,2H), 3.493 (t, J=6.0 Hz, 2H), 4.43 (s, 2H), 6.97 (s, 2H), 7.21 (d, J=8.0Hz, 1H), 7.88 (s, 1H), 7.90 (d, J=8.4 Hz, 1H).

General Amine Alkylation Procedure for the Synthesis of theAminoacetonitrile Precursors.

The aminoacetonitrile precursors are synthesized according to theprotocol described in Ruchelman, A. L.; Houghton, P. J.; Zhou, N.; Liu,A.; Liu, L. F.; LaVoie, E. J. 5-(2-Aminoethyl)dibenzo[c,h][1,6]naphthyridin-6-ones: Variation of N-Alkyl SubstituentsModulates Sensitivity to Efflux Transporters Associated with MultidrugResistance J. Med. Chem. 2005, 48, 792-804, incorporated herein byreference in its entirety for any and all purposes. Thus, theappropriate secondary amine is combined with chloroacetonitrile (1.1equiv.), K₂CO₃ (2.0 equiv.) and potassium iodide (1.0 equiv.) inacetonitrile (2.5 mL/mmol of secondary amine). The reaction is stirredat room temperature (“rt”, about 21° C.) for 13-16 h, diluted withaqueous saturated Na₂CO₃ (50 mL) and extracted with ether (3×30 mL). Thecombined organics were washed with brine and dried with Na₂SO₄. Thesolvent was removed in vacuo and the residue purified by silica gelchromatography to afford the necessary aminoacetonitrile precursors.Select representative synthesis of such aminoacetonitrile precursors areprovided in Examples 10-13 below for the synthesis of aminoacetonitrileprecursors to diamine fragments 4a-4d.

Example 10: 2-(Benzyl(tert-butyl)amino)acetonitrile

To a solution of N-benzyl-tert-butylamine (930 mg, 5.70 mmol) in MeCN(15 mL) was added K₂CO₃ (1,575 mg, 11.40 mmol, 2 equiv), potassiumiodide (946 mg, 5.70 mmol, 1 equiv) and chloroacetonitrile (1,720 mg,22.79 mmol, 4 equiv). The reaction was heated at 75° C. for 16 h, cooledto rt and partitioned between water (150 mL) and ethyl ether (3×75 mL).The combined organic layers were washed with brine (50 mL), dried(Na₂SO₄), adsorbed onto celite and purified by silica gel chromatographyto afford the nitrile product as a colorless oil (846 mg, 4.18 mmol, 73%yield). R_(f)=0.49 (10% EtOAc/hexanes); ¹H NMR (400 MHz, CDCl₃) δ 1.29(s, 9H), 3.43 (s, 2H), 3.82 (s, 2H), 7.23-7.28 (m, 1H), 7.29-7.36(complex, 4H); ¹³C NMR (101 MHz, CDCl₃, APT pulse sequence) δ d 27.4,127.5, 128.5, 128.6; u 35.7, 51.4, 55.2, 118.0, 138.9; IR 2975, 1454,1366, 1202 cm⁻¹; HRMS (ESI) m/z calcd for C₁₃H₁₉N₂ ([M+H]⁺) 203.1543,found 203.1536.

Example 11: 2-((4-Fluorobenzyl)(tert-butyl)amino)acetonitrile

N-(4-Fluoromobenzyl)-tert-butylamine (900 mg, 4.97 mmol) was reactedaccording to the protocol in Example 10 to afford the nitrile product asa colorless oil (1,038 mg, 4.71 mmol, 95% yield). R_(f)=0.49 (10%EtOAc/hexanes); ¹H NMR (400 MHz, CDCl₃) δ 1.29 (s, 9H), 3.42 (s, 2H),3.80 (s, 2H), 7.00 (t, J=8.8 Hz, 2H), 7.31 (dd, J=5.6, 8.8 Hz, 2H); ¹³CNMR (101 MHz, CDCl₃, APT pulse sequence) δ d 27.4, 115.5 (d, J=21.5 Hz),130.1 (d, J=8.0 Hz); u 35.6, 50.7, 55.3, 117.9, 134.5, 162.3 (d, J=246Hz); IR 2975, 1604, 1508, 1367, 1219 cm⁻¹; HRMS (ESI) m/z calcd forC₁₃H₁₈FN₂ ([M+H]⁺) 221.1449, found 221.1438.

Example 12: 2-((4-Chlorobenzyl)(tert-butyl)amino)acetonitrile

N-(4-Chlorobenzyl)-tert-butylamine (700 mg, 3.54 mmol) was reactedaccording to the protocol in Example 10 to afford the nitrile product asa white solid (685 mg, 2.89 mmol, 82% yield). Mp=65-67° C.; R_(f)=0.49(10% EtOAc/hexanes); ¹H NMR (400 MHz, CDCl₃) δ 1.27 (s, 9H), 3.41 (s,2H), 3.78 (s, 2H), 7.27 (s, 4H); ¹³C NMR (101 MHz, CDCl₃, APT pulsesequence) δ d 27.2, 128.6, 129.8; u 35.6, 50.7, 55.2, 117.7, 133.0,137.4; IR 2975, 1597, 1490, 1368, 1201 cm⁻¹; HRMS (ESI) m/z calcd forC₁₃H₁₈ClN₂ ([M+H]⁺) 237.1153, found 237.1141.

Example 13: 2-((4-Bromobenzyl)(tert-butyl)amino)acetonitrile

N-(4-Bromobenzyl)-tert-butylamine (920 mg, 3.80 mmol) was reactedaccording to the protocol in Example 10 to afford the nitrile product asa white solid (906 mg, 3.22 mmol, 85% yield). Mp=64-66° C.; R_(f)=0.49(10% EtOAc/hexanes); ¹H NMR (400 MHz, CDCl₃) δ 1.26 (s, 9H), 3.40 (s,2H), 3.76 (s, 2H), 7.21 (d, J=8.4 Hz, 2H), 7.42 (d, J=8.0 Hz, 2H); ¹³CNMR (101 MHz, CDCl₃, APT pulse sequence) δ d 27.2, 130.1, 131.5; u 35.6,50.7, 55.1, 117.6, 121.1, 137.9; IR 2976, 1592, 1485, 1366, 1203 cm⁻¹;HRMS (ESI) m/z calcd for C₁₃H₁₈BrN₂ ([M+H]⁺) 281.0648, found 281.0644.

General Reduction Procedure for the Synthesis of the Diamine Fragments.

The diamine fragments were synthesized according to the protocoldescribed in Ruchelman, A. L.; Houghton, P. J.; Zhou, N.; Liu, A.; Liu,L. F.; LaVoie, E. J. 5-(2-Aminoethyl)dibenzo[c,h][1,6]naphthyridin-6-ones: Variation of N-Alkyl SubstituentsModulates Sensitivity to Efflux Transporters Associated with MultidrugResistance J. Med. Chem. 2005, 48, 792-804, incorporated herein byreference in its entirety for any and all purposes. Thus, to a solutionof the appropriate aminoacetonitrile precursor in THF or ether was addedlithium aluminum hydride (3.5 M solution in THF, 1.1 equiv.). Thereaction was stirred for 13-16 h, carefully quenched with EtOAc thenwater, acidified with aqueous HCl (2 M) and extracted with EtOAc (3×30mL). The pH of the aqueous layer was adjusted to >8 with aqueous NaOH (2M) and extracted with EtOAc (3×30 mL). The combined organics were driedwith Na2SO4 and the solvent removed in vacuo to afford the diaminefragments which were used without further purification.

Example 14: Exemplary Diamine Synthesis ofN¹-(4-bromobenzyl)-N¹-isopropylethane-1,2-diamine

2-((4-bromobenzyl)(isopropyl)amino)acetonitrile (459 mg, 1.72 mmol) wasreacted according to the general reduction procedure described above inether to afford the diamine (384 mg, 1.42 mmol, 82% yield) as a lightyellow oil. R_(f)=0.62 (10% MeOH, 1% Et3N in CH2Cl2); ¹H NMR (400 MHz,CDCl₃) δ 1.01 (d, J=6.4 Hz, 6H), 2.47 (t, J=6.0 Hz, 2H), 2.61 (t, J=6.0Hz, 2H), 2.89 (sept, J=6.4 Hz, 1H), 3.50 (s, 2H), 7.21 (d, J=8.8 Hz,2H), 7.41 (d, J=8.4 Hz, 2H); ¹³C NMR (101 MHz, CDCl₃) δ d 17.8 (×2),49.8, 130.0 (×2), 131.1 (×2); u 40.2, 52.3, 53.7, 120.2, 140.3; IR(neat) 2963, 1485 cm⁻¹; HRMS (ESI) m/z calcd for C₁₂H₂₀BrN₂ ([M+H]⁺),271.0810, found 271.0827.

Representative Synthesis of Compounds of the Present Technology.

Representative syntheses are provided below. Notably, the synthesis ofcomparative compound 1a is also provided, where 1a was later assayedwith representative compounds of the present technology.

Example 15: Comparative compoundN-(2-(Benzyl(isopropyl)amino)ethyl)-N-methyl-4-((N,4-dimethylphenylsulfonamido)methyl)benzamide(1a)

To a solution ofN-(2-(benzyl(isopropyl)amino)ethyl)-4-(((4-methylphenyl)sulfonamido)methyl)benzamide (41 mg, 0.085 mmol) in DMF (1 mL) was added sodiumhydride, 60% dispersion in mineral oil (10 mg, 0.256 mmol, 3 equiv). Thereaction was stirred for 10 min at rt and methyl iodide (30 mg, 0.215mmol, 2.5 equiv) was added. The reaction was stirred at rt for 17 h andpartitioned between aqueous NaHCO₃ and CH₂Cl₂. The organics wereseparated and the aqueous layer extracted with CH₂Cl₂ (3×5 mL) and thecombined organic layers were dried (Na₂SO₄), concentrated and purifiedby silica gel chromatography to afford 1a as a light yellow oil (36 mg,0.071 mmol, 83% yield). R_(f)=0.49 (75% EtOAc/hexanes); ¹H NMR (400 MHz,CDCl₃) (ca. 1:1 mixture of rotomers) δ 0.92 (d, J=6.4 Hz, 3H), 1.06 (d,J=6.4 Hz, 3H), 2.46 (s, 3H), 2.50 (t, J=7.2 Hz, 1H), 2.54 (s, 1.5H),2.59 (s, 1.5H), 2.73 (m, 1.5H), 2.85 (s, 1.5H), 2.93 (s, 1.5H), 3.04 (m,0.5H), 4.14 (m, 1H), 3.41 (s, 1H), 3.49 (t, J=6.0 Hz, 1H), 3.63 (s, 1H),4.13 (d, J=3.6 Hz, 2H), 7.19-7.37 (complex, 11H), 7.73 (d, J=8.0 Hz,2H); ¹³C NMR (101 MHz, CDCl₃, APT pulse sequence) (mixture of rotomers)δ d 17.9, 21.5, 33.6, 34.3, 34.5, 38.6, 49.8, 50.5, 126.7, 126.9, 127.0,127.3, 127.5, 128.2, 128.4, 128.7, 129.8; u 46.9, 47.1, 47.9, 51.0,53.9, 54.6, 134.4, 136.4, 136.5, 136.9, 137.0, 140.4, 140.9, 143.6,170.7, 171.5; IR 2964, 2928, 1629 cm⁻¹; HRMS (ESI) m/z calcd forC₂₉H₃₈N₃O₃S ([M+H]⁺) 508.2628, found 508.2629; HPLC purity=97.4%.

Example 16:N-(2-(tert-butyl(4-methoxybenzyl)amino)ethyl)-2-tosyl-1,2,3,4-tetrahydroisoquinoline-6-carboxamide(1b)

To a solution of 2-tosyl-1,2,3,4-tetrahydroisoquinoline-6-carboxylicacid 2a (100 mg, 0.30 mmol), diamine fragment 4e (71 mg, 0.30 mmol) anddiisopropylethylamine (0.16 mL, 0.90 mmol, 3.0 equiv) in DMF (3 mL) wasadded HATU (125 mg, 0.33 mmol, 1.1 equiv). The reaction was stirred for18 h at rt and all solvents removed in vacuo. The residue was purifiedby mass-directed, reverse phase preparative HPLC to afford 1b (131 mg,0.24 mmol, 79% yield). ¹H NMR (400 MHz, CDCl₃) δ 1.15 (s, 9H), 2.42 (s,3H), 2.80 (dd, J=5.1, 6.6 Hz, 2H), 2.95 (t, J=5.9 Hz, 1H), 3.11-3.23 (m,2H), 3.36 (t, J=5.9 Hz, 2H), 3.61 (s, 2H), 3.61 (s, 3H), 4.26 (s, 2H),6.17 (t, J=5.3 Hz, 1H), 6.72-6.77 (m, 2H), 7.03 (d, J=8.0 Hz, 1H),7.20-7.25 (m, 3H), 7.31-7.37 (m, 3H), 7.67-7.80 (m, 2H); ¹³C NMR (101MHz, CDCl₃) δ 21.7, 27.5, 29.0, 40.2, 43.7, 47.7, 49.9, 54.7, 55.3,55.6, 114.0, 124.5, 126.5, 127.8, 127.9, 129.2, 129.9, 133.2, 133.5,134.6, 134.9, 144.0, 158.5, 166.8; HRMS (ESI) m/z calcd for C₃₁H₄₀N₃O₄S([M+H]⁺), 550.2740, found 550.2719; HPLC purity=97.2%.

Example 17:N-(2-(Benzyl(isopropyl)amino)ethyl)-2-(mesitylsulfonyl)-1,2,3,4-tetrahydroisoquinoline-6-carboxamide(1c)

To a solution of2-(mesitylsulfonyl)-1,2,3,4-tetrahydroisoquinoline-6-carbonyl chloride(20 mg, 0.054 mmol) in CH₂Cl₂ (3 mL) was added diamine fragment 3a (10mg, 0.054 mmol) and triethylamine (0.019 mL, 0.14 mmol, 2.6 equiv). Thereaction was stirred for 48 h at rt, aqueous, saturated sodiumbicarbonate solution (2 mL) was added and all solvents removed in vacuo.The residue was extracted with a solution of CH₂Cl₂:MeOH (5:1, 6 mL).The filtrate was dried (Na₂SO₄), evaporated and purified bymass-directed, reverse phase preparative HPLC to afford 1c (9 mg, 0.017mmol, 31% yield). ¹H NMR (400 MHz, CDCl₃) δ 1.00 (d, J=6.6 Hz, 6H), 2.24(s, 3H), 2.57 (s, 6H), 2.61 (t, J=5.8 Hz, 2H), 2.83 (t, J=5.9 Hz, 2H),2.94 (sep, J=6.6 Hz, 1H), 3.29 (q, J=5.0 Hz, 2H), 3.41 (t, J=5.9 Hz,2H), 3.49 (s, 2H), 4.33 (s, 2H), 6.47 (br s, 1H), 6.90 (s, 2H), 7.03 (d,J=8.0 Hz, 1H), 7.15-7.30 (m, 6H), 7.39 (s, 1H); ¹³C NMR (101 MHz, CDCl₃)δ 18.0, 21.0, 22.9, 28.7, 37.5, 41.9, 45.7, 47.8, 49.7, 53.7, 124.4,126.6, 127.0, 127.9, 128.5, 128.6, 131.6, 132.0, 133.3, 133.8, 135.4,140.6, 140.8, 142.9, 166.7; IR 3378, 2965, 1648, 1544 cm⁻¹; HRMS (ESI)m/z calcd for C₃₁H₃₉N₃O₃S ([M+H]⁺), 534.2790, found 534.2804; HPLCpurity=98.7%.

Example 18:N-(2-((4-Fluorobenzyl)(isopropyl)amino)ethyl)-2-tosyl-1,2,3,4-tetrahydroisoquinoline-6-carboxamide(1d)

2-Tosyl-1,2,3,4-tetrahydroisoquinoline-6-carbonyl chloride (20 mg, 0.054mmol) and diamine fragment 3b (11 mg, 0.054 mmol) were reacted accordingthe protocol in Example 17 to afford 1d (15 mg, 0.028 mmol, 52% yield).¹H NMR (400 MHz, DMSO-d₆) δ 0.96 (d, J=6.4 Hz, 6H), 2.39 (s, 3H), 2.51(m, 2H), 2.82-2.90 (m, 3H), 3.22 (m, 2H), 3.29 (t, J=6.4 Hz, 2H), 3.54(s, 2H), 4.21 (s, 2H), 7.06 (t, J=8.8 Hz, 2H), 7.23 (d, J=8.4 Hz, 1H),7.36 (dd, J=6.4, 8.8 Hz, 2H), 7.43 (d, J=8.0 Hz, 2H), 7.56 (m, 2H), 7.72(d, J=8.4 Hz, 2H); ¹³C NMR (101 MHz, DMSO-d₆) δ 17.9, 21.0, 28.0, 40.4,43.4, 47.2, 48.4, 49.3, 53.0, 114.7 (d, J=21.1 Hz), 124.7, 126.4,127.45, 127.49, 129.9 130.0, 132.90, 132.96, 132.99, 134.6, 137.0 (d,J=2.8 Hz), 143.7, 161.0 (d, J=242.3 Hz), 165.6; IR 2966, 1649, 1508cm⁻¹; HRMS (ESI) m/z calcd for C₂₉H₃₅FN₃O₃S ([M+H]⁺), 524.2383, found524.2401; HPLC purity=97.0%.

Example 19:N-(2-((4-Chlorobenzyl)(isopropyl)amino)ethyl)-2-tosyl-1,2,3,4-tetrahydroisoquinoline-6-carboxamide(1e)

2-Tosyl-1,2,3,4-tetrahydroisoquinoline-6-carbonyl chloride (20 mg, 0.054mmol) and diamine fragment 3c (12 mg, 0.054 mmol) were reacted accordingthe protocol in Example 17 to afford 1e (13 mg, 0.024 mmol, 45% yield).¹H NMR (400 MHz, DMSO-d₆) δ 0.95 (d, J=6.4 Hz, 6H), 2.31 (s, 3H), 2.55(m, 2H), 2.85 (m, 2H), 3.24-3.28 (m, 4H), 3.41 (s, 2H), 4.17 (s, 2H),6.97 (d, J=8.0 Hz, 1H), 7.07-7.12 (complex, 4H), 7.15 (s, 1H), 7.19 (m,1H), 7.22 (d, J=8.0 Hz, 2H), 7.31 (br s, 1H), 7.62 (d, J=8.4 Hz, 2H);¹³C NMR (101 MHz, DMSO-d₆) δ 17.9, 21.0, 28.0, 40.4, 43.4, 47.2, 48.6,49.5, 53.0, 124.7, 126.4, 127.4, 127.5, 127.9, 129.86, 129.95, 130.2,130.9, 132.9, 133.0, 134.6, 140.1 143.6, 165.6; IR 2966, 1647, 1543,1491 cm⁻¹; HRMS (ESI) m/z calcd for C₂₉H₃₅ClN₃O₃S ([M+H]⁺), 540.2088,found 540.2104; HPLC purity=100.0%.

Example 20:N-(2-((4-Bromobenzyl)(isopropyl)amino)ethyl)-2-tosyl-1,2,3,4-tetrahydroisoquinoline-6-carboxamide(1f)

2-Tosyl-1,2,3,4-tetrahydroisoquinoline-6-carbonyl chloride (20 mg, 0.054mmol) and diamine fragment 3d (15 mg, 0.054 mmol) were reacted accordingthe protocol in Example 17 to afford 1f (18 mg, 0.30 mmol, 56% yield).¹H NMR (400 MHz, CDCl₃) δ 0.98 (d, J=6.4 Hz, 6H), 2.35 (s, 3H), 2.58 (t,J=5.6 Hz, 2H), 2.88-2.94 (m, 3H), 3.26-3.31 (m, 4H), 3.42 (s, 2H), 4.21(s, 2H), 6.39 (br s, 1H), 7.01 (d, J=8.0 Hz, 1H), 7.08 (d, J=8.4 Hz,1H), 7.22-7.28 (complex, 5H), 7.34 (s, 1H), 7.66 (d, J=8.4 Hz, 2H); ¹³CNMR (101 MHz, CDCl₃) δ 18.1, 21.6, 29.0, 37.8, 43.7, 47.6, 48.2, 49.9,53.2, 120.7, 124.4, 126.6, 127.75, 127.82, 129.9, 130.4, 131.6, 133.2,133.3, 133.7, 135.1, 139.9 144.0, 166.9; IR 2965, 1646, 1541, 1486 cm⁻¹;HRMS (ESI) m/z calcd for C₂₉H₃₄BrN₃O₃S ([M+H]⁺), 586.1562, found586.1585; HPLC purity=94.6%.

Example 21:N-(2-((4-Bromobenzyl)(isopropyl)amino)ethyl)-2-((4-ethylphenyl)sulfonyl)-1,2,3,4-tetrahydroisoquinoline-6-carboxamide(1g)

2-((4-Ethylphenyl)sulfonyl)-1,2,3,4-tetrahydroisoquinoline-6-carbonylchloride (160 mg, 0.44 mmol) and diamine fragment 3d (119 mg, 0.44 mmol)were reacted according the protocol in Example 17 to afford 1g (122 mg,0.21 mmol, 47% yield). ¹H NMR (400 MHz, DMSO-d₆) δ 0.95 (d, J=6.8 Hz,6H), 1.15 (t, J=7.6 Hz, 3H), 2.55 (t, J=6.0 Hz, 2H), 2.61 (q, J=7.6 Hz,2H), 2.83-2.90 (m, 3H), 3.22-3.28 (m, 4H) 3.40 (s, 2H), 4.18 (s, 2H),6.52 (br s, 1H), 6.99 (d, J=8.0 Hz, 1H), 7.07 (d, J=8.4 Hz, 1H)7.21-7.27 (complex, 5H), 7.33 (s, 1H), 7.65 (d, J=8.4 Hz, 2H); ¹³C NMR(101 MHz, DMSO-d₆) δ 15.0, 17.8, 27.98, 28.03, 40.4, 43.4, 47.2, 48.6,49.5, 53.1, 119.3, 124.7, 126.3, 127.47, 127.55, 128.7, 130.3, 130.8,132.92, 132.93, 133.1, 134.6, 140.5 149.5, 165.6; IR 2965, 1646, 1541,1486 cm⁻¹; HRMS (ESI) m/z calcd for C₃₀H₃₆BrN₃O₃S ([M+H]⁺), 600.1719,found 600.1740; HPLC purity=98.1%.

Example 22:N-(2-(Benzyl(tert-butyl)amino)ethyl)-2-tosyl-1,2,3,4-tetrahydroisoquinoline-6-carboxamide(1h)

2-Tosyl-1,2,3,4-tetrahydroisoquinoline-6-carbonyl chloride (20 mg, 0.054mmol) and diamine fragment 4a (11 mg, 0.054 mmol) were reacted accordingthe protocol in Example 17 to afford 1h (26 mg, 0.049 mmol, 91% yield).¹H NMR (400 MHz, CDCl₃) δ 1.10 (s, 9H), 2.36 (s, 3H), 2.76 (t, J=6.0 Hz,2H), 2.89 (t, J=6.0 Hz, 2H), 3.12 (q, J=5.6 Hz, 2H), 3.30 (t, J=6.0 Hz,2H), 3.64 (s, 2H), 4.21 (s, 2H), 6.22 (br s, 1H), 6.98 (d, J=8.0 Hz,1H), 7.09 (m, 1H), 7.16-7.22 (m, 3H), 7.27-7.29 (complex, 5H), 7.66 (d,J=8.4 Hz, 2H); ¹³C NMR (101 MHz, CDCl₃) δ 21.6, 27.5, 28.9, 39.9, 43.7,47.6, 50.1, 55.2, 55.6, 124.5, 126.5, 126.8, 127.7, 127.8, 127.9, 128.5,129.9, 133.2, 133.36, 133.42, 134.9, 142.9, 143.9, 166.7; IR 2989, 1628,1538, 1497 cm⁻¹; HRMS (ESI) m/z calcd for C₃₀H₃₇N₃O₃S ([M+H]⁺),520.2634, found 520.2650; HPLC purity=97.5%.

Example 23:N-(2-(Benzyl(tert-butyl)amino)ethyl)-((4-ethylphenyl)sulfonyl)-1,2,3,4-tetrahydroisoquinoline-6-carboxamide(1i)

2-((4-Ethylphenyl)sulfonyl)-1,2,3,4-tetrahydroisoquinoline-6-carbonylchloride (160 mg, 0.44 mmol) and diamine fragment 4a (91 mg, 0.44 mmol)were reacted according the protocol in Example 17 to afford 1i (66 mg,0.12 mmol, 28% yield). ¹H NMR (400 MHz, CDCl₃) δ 1.11 (s, 9H), 1.20 (t,J=7.6 Hz, 3H), 2.67 (q, J=7.6 Hz, 2H), 2.77 (t, J=6.1 Hz, 2H), 2.90 (t,J=5.9 Hz, 2H), 3.12 (q, J=5.9 Hz, 2H), 3.31 (t, J=5.9 Hz, 2H), 3.65 (s,2H), 4.21 (s, 2H), 6.37 (br s, 1H), 7.00 (d, J=8.0 Hz, 1H), 7.07-7.11(m, 1H), 7.16-7.21 (m, 2H), 7.24-7.27 (m, 1H), 7.29-7.33 (m, 5H),7.67-7.71 (m, 2H); ¹³C NMR (101 MHz, CDCl₃) δ 15.0, 27.3, 28.7, 28.8,39.9, 40.9, 43.6, 47.5, 50.0, 55.1, 124.5, 126.4, 126.6, 127.6, 127.76,127.81, 128.3, 128.6, 133.1, 133.2, 134.7, 142.8, 149.9, 166.6; IR 2968,1645, 1540, 1494 cm⁻¹; HRMS (ESI) m/z calcd for C₃₁H₃₉N₃O₃S ([M+H]⁺),534.2790, found 534.2801; HPLC purity=94.7%.

Example 24:N-(2-((4-Fluorobenzyl)(tert-butyl)amino)ethyl)-2-tosyl-1,2,3,4-tetrahydroisoquinoline-6-carboxamide(1j)

2-Tosyl-1,2,3,4-tetrahydroisoquinoline-6-carbonyl chloride (20 mg, 0.054mmol) and diamine fragment 4b (12 mg, 0.054 mmol) were reacted accordingthe protocol in Example 17 to afford 1j (28 mg, 0.052 mmol, 96% yield).¹H NMR (400 MHz, CDCl₃) δ 1.08 (s, 9H), 2.35 (s, 3H), 2.73 (t, J=6.1 Hz,2H), 2.87 (t, J=5.9 Hz, 2H), 3.10 (q, J=5.6 Hz, 2H), 3.29 (t, J=5.9 Hz,2H), 3.58 (s, 2H), 4.20 (s, 2H), 6.10 (br s, 1H), 6.77-6.84 (m, 2H),6.97 (d, J=8.0 Hz, 1H), 7.18-7.28 (m, 6H), 7.64-7.67 (m, 2H); ¹³C NMR(101 MHz, CDCl₃) δ 21.5, 27.4, 28.8, 40.1, 43.6, 47.5, 50.0, 54.5, 55.5,115.2 (d, J=21.2 Hz), 124.3, 126.4, 127.6, 127.7, 129.3 (d, J=8.0 Hz),129.8, 133.2, 133.3, 133.5, 135.0, 138.3 (d, J=4.0 Hz), 143.9, 161.6 (d,J=245.4 Hz), 166.7; IR 2970, 1643, 1541, 1506 cm⁻¹; HRMS (ESI) m/z calcdfor C₃₀H₃₆FN₃O₃S ([M+H]⁺), 538.2540, found 538.2554; HPLC purity=96.3%.

Example 25:N-(2-((4-Fluorobenzyl)(tert-butyl)amino)ethyl)-2-(mesitylsulfonyl)-1,2,3,4-tetrahydroisoquinoline-6-carboxamide(1k)

2-(mesitylsulfonyl)-1,2,3,4-tetrahydroisoquinoline-6-carbonyl chloride(20 mg, 0.054 mmol) and diamine fragment 4b (12 mg, 0.054 mmol) werereacted according the protocol in Example 17 to afford 1k (14 mg, 0.024mmol, 45% yield). ¹H NMR (400 MHz, CDCl₃) δ 1.08 (s, 9H), 2.35 (s, 3H),2.73 (t, J=6.1 Hz, 2H), 2.87 (t, J=5.9 Hz, 2H), 3.10 (q, J=5.6 Hz, 2H),3.29 (t, J=5.9 Hz, 2H), 3.58 (s, 2H), 4.20 (s, 2H), 6.10 (br s, 1H),6.77-6.84 (m, 2H), 6.97 (d, J=8.0 Hz, 1H), 7.18-7.28 (m, 6H), 7.64-7.67(m, 2H); ¹³C NMR (101 MHz, CDCl₃) δ 21.5, 27.4, 28.8, 40.1, 43.6, 47.5,50.0, 54.5, 55.5, 115.2 (d, J=21.2 Hz), 124.3, 126.4, 127.6, 127.7,129.3 (d, J=8.0 Hz), 129.8, 133.2, 133.3, 133.5, 135.0, 138.3 (d, J=4.0Hz), 143.9, 161.6 (d, J=245.4 Hz), 166.7; IR 2971, 1644, 1604, 1543cm⁻¹; HRMS (ESI) m/z calcd for C₃₂H₄₁FN₃O₃S ([M+H]⁺), 566.2853, found566.2871; HPLC purity=99.1%.

Example 26:N-(2-((4-Chlorobenzyl)(tert-butyl)amino)ethyl)-2-tosyl-1,2,3,4-tetrahydroisoquinoline-6-carboxamide(1l)

2-Tosyl-1,2,3,4-tetrahydroisoquinoline-6-carbonyl chloride (20 mg, 0.054mmol) and diamine fragment 4c (13 mg, 0.054 mmol) were reacted accordingthe protocol in Example 17 to afford 1l (25 mg, 0.046 mmol, 85% yield).¹H NMR (400 MHz, CDCl₃) δ 1.09 (s, 9H), 2.39 (s, 3H), 2.64 (m, 2H), 2.87(t, J=6.0 Hz, 2H), 3.04 (m, 2H), 3.28 (t, J=6.0 Hz, 2H), 3.69 (s, 2H),4.20 (s, 2H), 7.20 (d, J=8.0 Hz, 1H), 7.31 (m, 2H), 7.39-7.45 (m, 4H),7.53 (m, 2H), 7.71 (d, J=8.4 Hz, 2H); ¹³C NMR (101 MHz, CDCl₃) δ 21.0,27.1, 28.0, 40.4, 43.4, 47.2, 49.9, 53.4, 54.8, 124.7, 126.3, 127.42,127.45, 127.8, 129.3, 129.9, 130.5, 132.77, 132.84, 132.9, 134.6, 142.1,143.6, 165.5; IR 2969, 1644, 1541 cm⁻¹; HRMS (ESI) m/z calcd forC₃₀H₃₇ClN₃O₃S ([M+H]⁺), 554.2244, found 554.2261; HPLC purity=99.2%.

Example 27:N-(2-((4-Bromobenzyl)(tert-butyl)amino)ethyl)-2-tosyl-1,2,3,4-tetrahydroisoquinoline-6-carboxamide(1m)

2-Tosyl-1,2,3,4-tetrahydroisoquinoline-6-carbonyl chloride (20 mg, 0.054mmol) and diamine fragment 4d (15 mg, 0.054 mmol) were reacted accordingthe protocol in Example 17 to afford 1m (7.8 mg, 0.013 mmol, 24% yield).¹H NMR (400 MHz, CDCl₃) δ 1.08 (s, 9H), 2.35 (s, 3H), 2.74 (t, J=6.0 Hz,2H), 2.89 (t, J=5.9 Hz, 2H), 3.13 (q, J=5.8 Hz, 2H), 3.27-3.32 (m, 2H),3.56 (s, 2H), 4.21 (s, 2H), 6.07 (br s, 1H), 7.00 (d, J=8.0 Hz, 1H),7.11-7.20 (m, 3H), 7.23-7.30 (m, 5H), 7.64-7.68 (m, 2H); ¹³C NMR (101MHz, CDCl₃) δ 21.6, 27.3, 28.9, 40.1, 43.6, 47.5, 50.2, 54.6, 55.6,113.9, 124.3, 126.5, 127.6, 127.7, 129.5, 129.8, 131.5, 133.19, 133.22,133.6, 135.0, 143.8, 166.7; IR 2980, 1652, 1521 cm⁻¹; HRMS (ESI) m/zcalcd for C₃₀H₃₆BrN₃O₃S ([M+H]⁺), 600.1719, found 600.1740; HPLCpurity=94.1%.

Example 28:N-(2-((4-Bromobenzyl)(tert-butyl)amino)ethyl)-2-(mesitylsulfonyl)-1,2,3,4-tetrahydroisoquinoline-6-carboxamide(1n)

2-(mesitylsulfonyl)-1,2,3,4-tetrahydroisoquinoline-6-carbonyl chloride(20 mg, 0.054 mmol) and diamine fragment 4d (15 mg, 0.054 mmol) werereacted according the protocol in Example 17 to afford 1n (10 mg, 0.016mmol, 29% yield). ¹H NMR (400 MHz, CDCl₃) δ 1.08 (s, 9H), 2.35 (s, 3H),2.74 (t, J=6.0 Hz, 2H), 2.89 (t, J=5.9 Hz, 2H), 3.13 (q, J=5.8 Hz, 2H),3.27-3.32 (m, 2H), 3.56 (s, 2H), 4.21 (s, 2H), 6.07 (br s, 1H), 7.00 (d,J=8.0 Hz, 1H), 7.11-7.20 (m, 3H), 7.23-7.30 (m, 5H), 7.64-7.68 (m, 2H);¹³C NMR (101 MHz, CDCl₃) δ 21.6, 27.3, 28.9, 40.1, 43.6, 47.5, 50.2,54.6, 55.6, 120.3, 124.3, 126.5, 127.6, 127.7, 129.5, 129.8, 131.5,133.21, 133.23, 133.6, 135.0, 141.9, 143.8, 166.7; IR 2972, 1647, 1533cm⁻¹; HRMS (ESI) m/z calcd for C₃₀H₃₆BrN₃O₃S ([M+H]⁺), 600.1719, found600.1740; HPLC purity=94.1%.

Representative Biological Activity of Compounds of the PresentTechnology

In Vitro Assay Methods

1. Compounds and Reagents

(+)-(5α,7α,8β)-N-Methyl-N-(7-(1-pyrrolidinyl)-1-oxaspiro(4.5)dec-8-yl)-benzeneacetamide(U69,593) and nor-binaltorphimine dihydrochloride (norBNI) werepurchased from Sigma Aldrich. The structure of U69,593 is providedbelow.

U69,593 was prepared in ethanol as a 10 mM stock, norBNI was prepared inwater as a 10 mM stock, and test compounds were prepared as 10 mM stocksin DMSO (Fisher). All compounds were then diluted to workingconcentrations in vehicle for each assay without exceeding 1% DMSO orethanol concentrations. [³⁵S]GTPγS was purchased from PerkinElmer LifeSciences. Phospho-ERK1/2 and total ERK1/2 antibodies were purchased fromCell Signaling (Beverly, Mass.) and Li-Cor secondary antibodies(anti-rabbit IRDye800CW and anti-mouse IRDye680LT) were purchased fromLi-Cor Biosciences.

2. Cell Lines and Cell Culture

Chinese hamster ovary (CHO) cells were virally transfected to expressHA-tagged recombinant human kappa opioid receptors (CHO-hKOR cell line)and maintained in DMEM/F-12 media (Invitrogen) supplemented with 10%fetal bovine serum, 1% penicillin/streptomycin, and 500 μg/ml geneticinas described in Schmid, C. L.; Streicher, J. M.; Groer, C. E.; Munro, T.A.; Zhou, L.; Bohn, L. M. Functional selectivity of6′-guanidinonaltrindole (6′-GNTI) at kappa opioid receptors in striatalneurons. J. Biol. Chem. 2013, 288, 22387-22398. A DiscoveRx PathHunter™U2OS cell line expressing βarrestin2 and hKOR (U2OS-hKOR-βarrestin2-DX)was purchased from DiscoveRx Corporation (Fremont, Calif.) andmaintained MEM with 10% fetal bovine serum, 1% penicillin/streptomycin,500 μg/ml geneticin and 250 μg/ml hygromycin B. All cells were grown at37° C. (5% CO₂ and 95% relative humidity).

3. Protein Coupling Assay

[³⁵S]GTPγS binding assay was performed following a previously publishedprotocol (Zhou, L.; Lovell, K. M.; Frankowski, K. J.; Slauson, S. R.;Phillips, A. M. Streicher, J. M.; Stahl, E.; Schmid, C. L.; Hodder, P.;Madoux, F.; Cameron, M. D.; Prisinzano, T. E.; Aubé, J.; Bohn, L. M. J.Biol. Chem. 2013, 288, 36703-36716). Briefly, cells were serum starvedfor 1 hour and membranes were prepared. Each reaction was performed atroom temperature and contained 15 μg of membrane protein, 40 μM GDP,˜0.1 nM [³⁵S]GTPγS and increasing concentrations of compounds in assaybuffer (50 mM Tris-HCl, pH 7.4, 100 mM NaCl, 5 mM MgCl₂, 1 mM EDTA).Directly after the addition of test compounds, 100 nM U69,593 was addedto yield a total volume of 200 μL. After 1 hour, reactions were quenchedby rapid filtration through GF/B filters and radioactivity was countedwith a TopCount NXT high throughput screening microplate scintillationand luminescence counter (PerkinElmer Life Sciences).

4. βArrestin2 Recruitment (DiscoveRx PathHunter™) Assay

The PathHunter™ assay was performed according to the manufacturer'sprotocol with slight modification and following known protocols (Schmid,C. L.; Streicher, J. M.; Groer, C. E.; Munro, T. A.; Zhou, L.; Bohn, L.M. Functional selectivity of 6′-guanidinonaltrindole (6′-GNTI) at kappaopioid receptors in striatal neurons. J. Biol. Chem. 2013, 288,22387-22398; Zhou, L.; Lovell, K. M.; Frankowski, K. J.; Slauson, S. R.;Phillips, A. M. Streicher, J. M.; Stahl, E.; Schmid, C. L.; Hodder, P.;Madoux, F.; Cameron, M. D.; Prisinzano, T. E.; Aubé, J.; Bohn, L. M.Development of functionally selective, small molecule agonists at kappaopioid receptors. J. Biol. Chem. 2013, 288, 36703-36716). Briefly, 5,000cells/well were plated overnight in Opti-MEM containing 1% fetal bovineserum, 1% penicillin/streptomycin. Cells were pretreated with antagonistfor 15 min at 37° C. followed by the addition of 1 μM U69,593 and a 90minute incubation at 37° C. PathHunter™ detection reagent was added andcells incubated at room temperature for 60 minutes. Chemiluminescencewas detected using a SpectraMax® M5e Multimode Plate Reader (MolecularDevices). βarrestin2 recruitment assays were also performed using animaging platform (Cellomics) to visualize translocation of a fluorescentprotein tagged parrestin2 to the membrane of KOR expressing U2OS cells.

5. In-Cell Western ERK1/2 Phosphorylation

Antagonist inhibition of U69,593 induced ERK phosphorylation wasdetermined by in-cell westerns via known protocols (Schmid, C. L.;Streicher, J. M.; Groer, C. E.; Munro, T. A.; Zhou, L.; Bohn, L. M.Functional selectivity of 6′-guanidinonaltrindole (6′-GNTI) at kappaopioid receptors in striatal neurons. J. Biol. Chem. 2013, 288,22387-22398; Zhou, L.; Lovell, K. M.; Frankowski, K. J.; Slauson, S. R.;Phillips, A. M. Streicher, J. M.; Stahl, E.; Schmid, C. L.; Hodder, P.;Madoux, F.; Cameron, M. D.; Prisinzano, T. E.; Aubé, J.; Bohn, L. M.Development of functionally selective, small molecule agonists at kappaopioid receptors. J. Biol. Chem. 2013, 288, 36703-36716). Briefly,hKOR-CHO cells were plated in 384-well plate at 15,000 cells per welland incubated at 37° C. overnight. After an hour serum starve, cellswere treated with compound followed by the addition of 100 nM U69,593and a 10 minute incubation at 37° C. Cells were fixed, permeabilized,blocked, and stained with primary antibodies for phosphorylated ERK1/2and total-ERK1/2 (1:300 and 1:400, respectively) at 4° C. overnight.Cells then incubated with Li-Cor secondary antibodies (anti-rabbitIRDye800CW, 1:500; anti-mouse IRDye680LT, 1:1500) and imaged with theOdyssey Infrared Imager (Li-Cor Biosciences, Lincoln, Nebr.) at 700 and800 nm.

6. Data Analysis and Statistics

GraphPad Prism 6.01 software (GraphPad) was used to generate sigmoidaldose response curves using a three-parameter, non-linear regressionanalysis. All compounds were run in parallel assays in 2-4 replicatesper individual experiment. All studies were performed n>3 independentexperiments in multiple replicates. For determination of antagonistinhibition, each individual experiment was normalized to the percentageof maximal U69,593 stimulation. The efficacy and potency values wereobtained from the averages of the nonlinear regression analysisperformed on each individual curve and are reported as the mean±S.E.M.

TABLE 1 entry/

IC₅₀ (nM) ± S.E.M.^(a) cmpd R¹ R² R³ [³⁵S]GTPγS ERK βarr2 norBNI — — —0.28 ± 0.03 4.6 ± 0.7 2.5 ± 0.3 ML140

138 ± 54  591 ± 87  403 1a

inactive inactive inactive 1c 2,4,6- i-Pr H 113 ± 42  1,120 ± 110   930± 150 Me 1d 4-Me i-Pr F 29.1 ± 7.9  406 ± 80  478 ± 110 1e 4-Me i-Pr Cl10.0 ± 1.4  400 ± 87  392 ± 91  1f 4-Me i-Pr Br 19.7 ± 6.6  258 ± 93 388 ± 85  1g 4-Et i-Pr Br 33 ± 11 314 ± 56  350 ± 68  1h 4-Me tert-Bu H5.28 ± 0.60 49.1 ± 9.7  103.7 ± 8.2  1i 4-Et tert-Bu H 3.41 ± 0.40 143 ±23  186 ± 57  1j 4-Me tert-Bu F 3.7 ± 1.2 76 ± 24 104 ± 32  1k 2,4,6-tert-Bu F 85 ± 20 1,200 ± 300   880 ± 67  Me 1l 4-Me tert-Bu Cl 1.6 ±0.5 107 ± 17  84 ± 20 1m 4-Me tert-Bu Br 2.9 ± 1.2 87 ± 17 93 ± 30 1n2,4,6- tert-Bu Br 106 ± 16  1,340 ± 410   710 ± 150 Me ^(a)n ≥ 3. Allcompounds fully blocked U69,593 (>86%)

Notably, all the compounds of the present technology (1c to 1n) weresignificantly more potent than ML140 in inhibiting G protein function.The effect of changing from isopropyl to tert-butyl was even morepronounced, providing compounds possessing single-digit nanomolarpotency. Halogen incorporation further improved the potency, resultingin the nearly equipotent fluoro analogue 1j, chloro analogue 1l, and thecorresponding bromide 1m. While 2,4,6-trimethyl aryl sulfonamidesubstitution led to notably less potent compounds when compared toeither the benzene sulfonamide, 4-methyl- or 4-ethyl-substituted arylsulfonamide analogues of the present technology, such compounds arestill more potent than ML140.

Table 2 provides the KOR recruitment of β arrestin 2 (percent response)toward a set concentration of the KOR agonist standard, U69,593.

TABLE 2 entry/

β arrestin 2 Inhibition (% U69,593 Stimulation) cmpd R¹ R² R³ 1 μM 100nM 10 nM norBNI — — — 0.9 0.6 0.1 1b 4-Me tert-Bu OMe 0.0 0.8 0.0 1l4-Me tert-Bu Cl 1.5 0.4 29.9

FIGS. 1A-D summarize the results of experiments comparing the detectedconcentrations of 1l (Example 26; referred to at AN4-015 in the Figure)or norBNI in the plasma and brain tissue of mice over the time periodnecessary for clearance (or up to 72 h) following a single 10 mg/kg IPdose in adult male C57Bl/6 mice according to methods discussed in Zhou,L.; Lovell, K. M.; Frankowski, K. J.; Slauson, S. R.; Phillips, A. M.Streicher, J. M.; Stahl, E.; Schmid, C. L.; Hodder, P.; Madoux, F.;Cameron, M. D.; Prisinzano, T. E.; Aubé, J.; Bohn, L. M. J. Biol. Chem.2013, 288, 36703-36716. Briefly, test compounds were dissolved anddelivered in 1:1:8 DMSO, Tween80, H₂O vehicle by IP injection. Plasmasamples were collected at the times indicated from the same cohort ofmice; brains were collected once at the times indicated. Plasma andbrain were mixed with acetonitrile (1:5 v:v or 1:5 w:v, respectively andthe brains were disrupted via sonication. Following centrifugation at16,000×g, the concentration of compound in the supernatant wasdetermined using liquid chromatography (Shimadzu, Japan)/tandem massspectrometry (AB Sciex, Franmingham, Mass.) operated in positive ionmode using multiple reaction monitoring methods. Separate standardcurves were prepared in blank plasma and brain matrix. For brain, theconcentration was calculated as amount of compound per mg tissue andconverted to a concentration assuming a density of 1 wherein 1 g oftissue equals 1 ml.

Compound 1l was found to cross the blood brain barrier into the CNS atconcentrations similar to norBNI within 0.5 h (FIGS. 1A & 1B). Moreover,1l was almost fully cleared from brain tissue by the 4 h measurement. Incontrast, norBNI was still significantly present after 72 h. The plasmalevel measurements followed a similar though less striking pattern(FIGS. 1C & 1D). These experiments demonstrate three key attributes ofthe compounds of the present technology: (1) the ability to cross theblood brain barrier and access the CNS, (2) attain therapeuticconcentrations in the brain tissue rapidly (within 1 h), and (3) thecompounds of the present technology are steadily removed from both theplasma and brain. Such a pharmacokinetic profile illustrates thesurprising and unexpected advantage of using compounds of the presenttechnology as short-acting KOR antagonists.

Assessment of KOR Mediated Physiological Responses:

Compounds of the present technology will be tested, on their own and/orin combination with a known KOR agonist, to assess the modulation ofphysiological responses including: antinociception, locomotor activity,pruritis and alterations in reward thresholds.

1. Antinociception

Studies will be performed using mouse nociceptive assays including thewarm water tail flick assay as known to those of ordinary skill in theart, such as discussed in Bohn L M, Lefkowitz R J, Caron M G.Differential mechanisms of morphine antinociceptive tolerance revealedin (beta)arrestin-2 knock-out mice. J Neurosci. 2002, 22(23):10494-500.Other tests of nociception include chemical-induced inflammatory painand visceral pain, as exemplified in Tarselli M. A., et al. Synthesis ofconolidine, a potent non-opiod analgesic for tonic and persistent pain.Nat. Chem. 2011, 3(6): 449-53. Antagonistic properties will be assessedby the compound's ability to block U50,488-induced antinociception. Thestructure of U50,488 is provided below.

2. Antipruritic Effects

Antipruritic effects will be determined by pretreating mice with the KORmodulator (i.e., a compound of the present technology or a known KORagonist). For agonists, this will be followed by a 40 mg/kg s.c. (napeof neck) injection of the pruritic agent, chloroquine phosphate and thenumber of scratching bouts in a 60 minute period will be counted, suchas described in as previously described Morgenweck, J., Frankowski, K.J., Prisinzano, T. E., Aubé, J., Bohn, L. M. Investigation of the roleof βarrestin2 in kappa opioid receptor modulation in a mouse model ofpruritus. Neuropharmacology, 2015, 99:600-609. For suspected negativemodulators, antagonism will be assessed by pretreating with a compoundof the present technology, followed by KOR agonist (U50,488H) then thepruritic agent. It is expected the compounds of the present technologywill block the antipruritic effect of U50,488 thus further demonstratingthat the compounds of the present technology display antagonism towardsnonbiased KOR agonists.

3. Locomotor Activity

Locomotor activity will be assessed by treating non-habituated mice (toreveal a measurable baseline activity as the reference KOR agonists willdecrease activity) prior to placing in open field testing boxes(Versamax by Accuscan Instruments), such as described in Morgenweck, J.F. K., Prisinzano, T. E., Aubé, J., Bohn, L. M. Investigation of therole of βarrestin2 in kappa opioid receptor modulation in a mouse modelof pruritus. Neuropharmacology, 2015, 99:600-609; Raehal, K. M., Schmid,C. L., Medvedev, I. O., Gainetdinov, R. R., Premont, R. T., Bohn, L. M.Morphine-induced physiological and behavioral responses in mice lackingG protein-coupled receptor kinase 6. Drug and alcohol dependence. 2009,104(3):187-96; Bohn, L. M., Gainetdinov, R. R., Sotnikova, T. D.,Medvedev, I. O., Lefkowitz, R. J., Dykstra, L. A., Caron, M. G. Enhancedrewarding properties of morphine, but not cocaine, in beta(arrestin)-2knock-out mice. The Journal of Neuroscience: the Official Journal of theSociety for Neuroscience, 2003, 23(32):10265-73; Medvedev, I. O.,Gainetdinov, R. R., Sotnikova, T. D., Bohn, L. M., Caron, M. G.,Dykstra, L. A. Characterization of conditioned place preference tococaine in congenic dopamine transporter knockout female mice.Psychopharmacology. 2005; 180(3):408-13; and Bohn, L. M., Xu, F.,Gainetdinov, R. R., Caron, M. G. Potentiated opioid analgesia innorepinephrine transporter knock-out mice. The Journal of Neuroscience:the Official Journal of the Society for Neuroscience, 2000,20(24):9040-5.

For example, adult male C57BL/6 mice from Jackson Labs (˜10 weeks old)were pretreated with either with vehicle (1:1:8, DMSO, Tween-80, dH₂O)orN-(2-((4-chlorobenzyl)(tert-butyl)amino)ethyl)-2-tosyl-1,2,3,4-tetrahydroisoquinoline-6-carboxamide(1l (Example 26) referred to at AN4015 in the Figure, 10 mg/kg, i.p.)and placed back in their home cage for 10 min. They were then treatedwith vehicle or U50,488 (10 l/g, s.c.) and placed in the open fieldactivity box (Versascan Instruments) for immediate recordings.Ambulatory behavior was recorded for 1 hour in 5 minute bins viadetection of infrared beam breaks. As shown in FIG. 2, Compound 1l wasobserved not to suppress activity compared to vehicle while U50, 488does (p<0.0001, U50,488 vs. vehicle; Two-way ANOVA for drug effect). Asillustrated in FIG. 3, a 10-minute pretreatment with Compound 1l (10mg/kg) prevents the development of U50,488 hypolocomotion compared tomice that received a vehicle pretreatment followed by U50,488 (p<0.0001,Two-way ANOVA for Compound 1l effect vs. U50,488 treatment). The animalnumbers are shown in parentheses in the legends of FIGS. 2 & 3.

4. Stress-Induced Reinstatement of Cocaine Conditioned Place Preference

Stress-induced reinstatement of cocaine conditioned place preferencestudies will be performed in C57Bl/6 mice, as discussed regardingoptimized cocaine-induced CPP in this strain as well as in lines derivedin part from this strain in Raehal, K. M., Schmid, C. L., Medvedev, I.O., Gainetdinov, R. R., Premont, R. T., Bohn, L. M. Morphine-inducedphysiological and behavioral responses in mice lacking G protein-coupledreceptor kinase 6. Drug Alcohol Depend. 2009, 104(3): 187-96; Medvedev,I. O., Gainetdinov, R. R., Sotnikova, T. D., Bohn, L. M., Caron, M. G.,Dykstra, L. A. Characterization of conditioned place preference tococaine in congenic dopamine transporter knockout female mice.Psychopharmacology (Berl). 2005, 180(3):408-13; and Bohn, L. M.,Gainetdinov, R. R., Sotnikova, T. D., Medvedev, I. O., Lefkowitz, R. J.,Dykstra, L. A., Caron, M. G. Enhanced rewarding properties of morphine,but not cocaine, in beta(arrestin)-2 knock-out mice. J Neurosci. 2003,23(32):10265-73. KOR antagonists have shown efficacious in preventingreinstatement in this line. A three department choice apparatus withambient lighting, wall color and floor texture serving as conditioningcues. A non-biased approach will be taken and only mice that do not showpreference during the preconditioning trials will be used. After cocainepreference is established, mice will be subjected to 7 days of dailysaline injection of only saline paired with each side of theconditioning chamber, to extinguish the association of the pairedchamber with the treatment of cocaine (10 mg/kg, i.p.). Extinction willbe verified by the loss of preference between the two chambers.Twenty-four hours post-extinction, mice will be injected with compoundsof the present technology (1-10 mg/kg, range determined based on PK andin vitro potency data) 20 minutes prior to being subjected to a stressorevent (a 6 min cold water swim) and then placed immediately in thepreference chambers. See Kreibich, A. S., Blendy, J. A. cAMP responseelement-binding protein is required for stress but not cocaine-inducedreinstatement. J Neurosci. 2004, 24(30):6686-92. Demonstration ofreinstatement will be an increase in time spent in the formerly cocainepaired side. U50,488 and/or Nor-BNI will serve as positive controls foreach paradigm.

While certain embodiments have been illustrated and described, a personwith ordinary skill in the art, after reading the foregoingspecification, can effect changes, substitutions of equivalents andother types of alterations to the compounds of the present technology orsalts, pharmaceutical compositions, derivatives, prodrugs, metabolites,tautomers or racemic mixtures thereof as set forth herein. Each aspectand embodiment described above can also have included or incorporatedtherewith such variations or aspects as disclosed in regard to any orall of the other aspects and embodiments.

The present technology is also not to be limited in terms of theparticular aspects described herein, which are intended as singleillustrations of individual aspects of the present technology. Manymodifications and variations of this present technology can be madewithout departing from its spirit and scope, as will be apparent tothose skilled in the art. Functionally equivalent methods within thescope of the present technology, in addition to those enumerated herein,will be apparent to those skilled in the art from the foregoingdescriptions. Such modifications and variations are intended to fallwithin the scope of the appended claims. It is to be understood thatthis present technology is not limited to particular methods, reagents,compounds, compositions, labeled compounds or biological systems, whichcan, of course, vary. It is also to be understood that the terminologyused herein is for the purpose of describing particular aspects only,and is not intended to be limiting. Thus, it is intended that thespecification be considered as exemplary only with the breadth, scopeand spirit of the present technology indicated only by the appendedclaims, definitions therein and any equivalents thereof.

The embodiments, illustratively described herein may suitably bepracticed in the absence of any element or elements, limitation orlimitations, not specifically disclosed herein. Thus, for example, theterms “comprising,” “including,” “containing,” etc. shall be readexpansively and without limitation. Additionally, the terms andexpressions employed herein have been used as terms of description andnot of limitation, and there is no intention in the use of such termsand expressions of excluding any equivalents of the features shown anddescribed or portions thereof, but it is recognized that variousmodifications are possible within the scope of the claimed technology.Additionally, the phrase “consisting essentially of” will be understoodto include those elements specifically recited and those additionalelements that do not materially affect the basic and novelcharacteristics of the claimed technology. The phrase “consisting of”excludes any element not specified.

In addition, where features or aspects of the disclosure are describedin terms of Markush groups, those skilled in the art will recognize thatthe disclosure is also thereby described in terms of any individualmember or subgroup of members of the Markush group. Each of the narrowerspecies and subgeneric groupings falling within the generic disclosurealso form part of the invention. This includes the generic descriptionof the invention with a proviso or negative limitation removing anysubject matter from the genus, regardless of whether or not the excisedmaterial is specifically recited herein.

As will be understood by one skilled in the art, for any and allpurposes, particularly in terms of providing a written description, allranges disclosed herein also encompass any and all possible subrangesand combinations of subranges thereof. Any listed range can be easilyrecognized as sufficiently describing and enabling the same range beingbroken down into at least equal halves, thirds, quarters, fifths,tenths, etc. As a non-limiting example, each range discussed herein canbe readily broken down into a lower third, middle third and upper third,etc. As will also be understood by one skilled in the art all languagesuch as “up to,” “at least,” “greater than,” “less than,” and the like,include the number recited and refer to ranges which can be subsequentlybroken down into subranges as discussed above. Finally, as will beunderstood by one skilled in the art, a range includes each individualmember.

All publications, patent applications, issued patents, and otherdocuments (for example, journals, articles and/or textbooks) referred toin this specification are herein incorporated by reference as if eachindividual publication, patent application, issued patent, or otherdocument was specifically and individually indicated to be incorporatedby reference in its entirety. Definitions that are contained in textincorporated by reference are excluded to the extent that theycontradict definitions in this disclosure.

Other embodiments are set forth in the following claims, along with thefull scope of equivalents to which such claims are entitled.

What is claimed is:
 1. A compound according to formula I

or stereoisomers, tautomers, solvates, and/or salts thereof; wherein G¹and G² are each independently C═O or S(O)₂; R¹, R², R³, R⁴, R⁵, R⁷, R⁸,R⁹, R¹⁰, and R¹¹ are each independently H, halo, hydroxy, amino, cyano,trifluoromethyl, thiol, alkylthio, sulfoxide, sulfone, nitro,pentafluorosulfanyl, carboxylate, amide, ester, or a substituted orunsubstituted C₁-C₆ alkyl, C₁-C₆ alkoxy, aryl, aryloxy, C₁-C₆ alkanoyl,C₁-C₈ alkanoyloxy, aryloyl, or aryloyloxy group, where any two adjacentR¹, R², R³, R⁴, R⁵, R⁷, R⁸, R⁹, R¹⁰, and R¹¹ may join to form a5-membered or 6-membered substituted or unsubstituted heteroalkyl group;R⁶ is a branched C₁-C₈ alkyl group or a substituted or unsubstitutedcycloalkyl or aryl group; and R¹² and R¹³ are each independently H or asubstituted or unsubstituted C₁-C₈ alkyl or C₅-C₇ cycloalkyl group; andn is 0, 1, or 2; provided that when G¹ is S(O)₂, G² is C═O, R¹, R², R⁴,R⁵, R⁷, R⁸, R⁹, R¹⁰, R¹¹, R¹², R¹³ are each H, R⁶ is isopropyl, and n is1, then R³ is not methyl.
 2. The compound of claim 1, wherein at leastone of R¹, R², R³, R⁴, R⁵, R⁷, R⁸, R⁹, R¹⁰, and R¹¹ is halo, hydroxy,cyano, trifluoromethyl, thiol, alkylthio, nitro, pentafluorosulfanyl,carboxylate, ester, or a substituted or unsubstituted C₁-C₆ alkyl, C₁-C₆alkoxy, aryl, aryloxy, C₁-C₆ alkanoyl, C₁-C₆ alkanoyloxy, aryloyl, oraryloyloxy group.
 3. The compound of claim 1, wherein at least one ofR¹, R², R³, R⁴, R⁵, R⁷, R⁸, R⁹, R¹⁰, and R¹¹ is hydroxy or a substitutedor unsubstituted C₁-C₆ alkoxy or C₁-C₆ alkanoyloxy group.
 4. Thecompound of claim 1, wherein at least one of R¹, R², R³, R⁴, R⁵, R⁷, R⁸,R⁹, R¹⁰, and R¹¹ is hydroxy or an unsubstituted C₁-C₆ alkoxy group. 5.The compound of claim 1, wherein at least one of R¹, R², R³, R⁴, and R⁵is halo, hydroxy, carboxylate, ester, or a substituted or unsubstitutedC₁-C₆ alkyl, C₁-C₆ alkoxy, C₁-C₆ alkanoyl, or C₁-C₆ alkanoyloxy group.6. The compound of claim 1, wherein at least one of R⁷, R⁸, R⁹, R¹⁰, andR¹¹ is halo, hydroxy, cyano, trifluoromethyl, thiol, alkylthio,sulfoxide, sulfone, nitro, pentafluorosulfanyl, carboxylate, ester, or asubstituted or unsubstituted C₁-C₆ alkyl, C₁-C₆ alkoxy, aryl, aryloxy,C₁-C₆ alkanoyl, C₁-C₆ alkanoyloxy, aryloyl, or aryloyloxy group.
 7. Thecompound of claim 1, wherein at least one of R⁷, R⁸, R⁹, R¹⁰, and R¹¹ ishydroxy or an unsubstituted C₁-C₆ alkoxy group.
 8. The compound of claim1, wherein at least two of R¹, R², R³, R⁴, R⁵, R⁷, R⁸, R⁹, R¹⁰, and R¹¹are each independently halo, hydroxy, cyano, trifluoromethyl, nitro,pentafluorosulfanyl, carboxylate, ester, or a substituted orunsubstituted C₁-C₆ alkyl, C₁-C₆ alkoxy, aryl, aryloxy, C₁-C₆ alkanoyl,C₁-C₆ alkanoyloxy, aryloyl, or aryloyloxy group.
 9. The compound ofclaim 1, wherein G¹ is S(O)₂; G² is C═O; R¹, R⁴, R⁵, R⁷, R¹⁰, and R¹¹are each H; R², R³, R⁸, and R⁹ are each independently halo, hydroxy,amino, cyano, trifluoromethyl, thiol, alkylthio, sulfoxide, sulfone,nitro, pentafluorosulfanyl, carboxylate, amide, ester, or a substitutedor unsubstituted C₁-C₆ alkyl, C₁-C₆ alkoxy, aryl, aryloxy, C₁-C₆alkanoyl, C₁-C₈ alkanoyloxy, aryloyl, or aryloyloxy group, where any twoadjacent R², R³, R⁸, and R⁹ may join to form a 5-membered or 6-memberedsubstituted or unsubstituted heteroalkyl group; R⁶ is a branched C₁-C₈alkyl group; R¹² and R¹³ are each H; and n is
 1. 10. The compound ofclaim 1, wherein G¹ is S(O)₂; G² is C═O; R¹, R⁴, R⁵, R⁷, R¹⁰, and R¹¹are each H; one of R² and R³ is halo, hydroxy, or a unsubstituted C₁-C₆alkyl, C₁-C₆alkoxy, C₁-C₆ alkanoyl, or C₁-C₈ alkanoyloxy group and theother R² or R³ is H; one of R⁸ and R⁹ is halo, hydroxy, amino, cyano,trifluoromethyl, thiol, alkylthio, sulfoxide, sulfone, nitro,pentafluorosulfanyl, carboxylate, amide, ester, or a substituted orunsubstituted C₁-C₆ alkoxy, C₁-C₆ alkanoyl, or C₁-C₈ alkanoyloxy group,and the other R⁸ or R⁹ is H; R⁶ is a branched C₁-C₈ alkyl group; R¹² andR¹³ are each H; and n is
 1. 11. The compound of claim 1, wherein R⁶ is abranched C₁-C₈ alkyl group or a substituted or unsubstituted cycloalkylgroup.
 12. The compound of claim 1, wherein R⁶ is isopropyl, sec-butyl,tert-butyl, isopentyl, neopentyl, or adamantyl.
 13. The compound ofclaim 1, wherein R⁶ is tert-butyl, neopentyl, or adamantyl.
 14. Thecompound of claim 1, wherein the compound is selected from the groupconsisting of


15. A composition comprising the compound of claim 1 and apharmaceutically acceptable carrier.
 16. A method comprising inhibitingβ-arrestin recruitment in a subject by administering an effective amountof a compound of claim
 1. 17. The method of claim 16, wherein thesubject is suffering from depression, alcohol addiction, or cocaineaddiction.
 18. A method of inhibiting β-arrestin recruitment wherein themethod comprises contacting a kappa opioid receptor with a compound ofclaim 1.